Ecor 1606 Assignment 1 Solutions Llc

  • [0001]

    The Sequence Listing associated with this application is provided on CD-ROM in lieu of a paper copy, and is hereby incorporated by reference into the specification. Three CD-ROMs are provided, containing identical copies of the sequence listing: CD-ROM No. 1 is labeled COPY 1, contains the file 563.app which is 2.2 MB and created on Feb. 1, 2002; CD-ROM No.2 is labeled COPY 2, contains the file 563.app which is 2.2 MB and created on Feb. 1, 2002; CD-ROM No. 3 is labeled CRF, contains the file 563.app which is 2.2 MB and created on Feb. 1, 2002.

  • [0002]

    1. Field of the Invention

  • [0003]

    The present invention relates generally to therapy and diagnosis of cancer, such as colon cancer. The invention is more specifically related to polypeptides, comprising at least a portion of a colon tumor protein, and to polynucleotides encoding such polypeptides. Such polypeptides and polynucleotides are useful in pharmaceutical compositions, e.g., vaccines, and other compositions for the diagnosis and treatment of colon cancer.

  • [0004]

    2. Description of the Related Art

  • [0005]

    Cancer is a significant health problem throughout the world. Although advances have been made in detection and therapy of cancer, no vaccine or other universally successful method for prevention and/or treatment is currently available. Current therapies, which are generally based on a combination of chemotherapy or surgery and radiation, continue to prove inadequate in many patients.

  • [0006]

    Colon cancer is the second most frequently diagnosed malignancy in the United States as well as the second most common cause of cancer death. The five-year survival rate for patients with colorectal cancer detected in an early localized stage is 92%; unfortunately, only 37% of colorectal cancer is diagnosed at this stage. The survival rate drops to 64% if the cancer is allowed to spread to adjacent organs or lymph nodes, and to 7% in patients with distant metastases.

  • [0007]

    The prognosis of colon cancer is directly related to the degree of penetration of the tumor through the bowel wall and the presence or absence of nodal involvement, consequently, early detection and treatment are especially important. Currently, diagnosis is aided by the use of screening assays for fecal occult blood, sigmoidoscopy, colonoscopy and double contrast barium enemas. Treatment regimens are determined by the type and stage of the cancer, and include surgery, radiation therapy and/or chemotherapy. Recurrence following surgery (the most common form of therapy) is a major problem and is often the ultimate cause of death. In spite of considerable research into therapies for the disease, colon cancer remains difficult to diagnose and treat. In spite of considerable research into therapies for these and other cancers, colon cancer remains difficult to diagnose and treat effectively. Accordingly, there is a need in the art for improved methods for detecting and treating such cancers. The present invention fulfills these needs and further provides other related advantages.

  • [0008]

    In spite of considerable research into therapies for these and other cancers, colon cancer remains difficult to diagnose and treat effectively. Accordingly, there is a need in the art for improved methods for detecting and treating such cancers. The present invention fulfills these needs and further provides other related advantages.

  • [0009]

    In one aspect, the present invention provides polynucleotide compositions comprising a sequence selected from the group consisting of:

  • [0010]

    (a) sequences provided in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417;

  • [0011]

    (b) complements of the sequences provided in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417;

  • [0012]

    (c) sequences consisting of at least 20, 25, 30, 35, 40, 45, 50, 75 and 100 contiguous residues of a sequence provided in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417;

  • [0013]

    (d) sequences that hybridize to a sequence provided in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417, under moderate or highly stringent conditions;

  • [0014]

    (e) sequences having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to a sequence of SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417;

  • [0015]

    (f) degenerate variants of a sequence provided in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417.

  • [0016]

    In one preferred embodiment, the polynucleotide compositions of the invention are expressed in at least about 20%, more preferably in at least about 30%, and most preferably in at least about 50% of colon tumor samples tested, at a level that is at least about 2-fold, preferably at least about 5-fold, and most preferably at least about 10-fold higher than that for normal tissues.

  • [0017]

    The present invention, in another aspect, provides polypeptide compositions comprising an amino acid sequence that is encoded by a polynucleotide sequence described above.

  • [0018]

    The present invention further provides polypeptide compositions comprising an amino acid sequence selected from the group consisting of sequences recited in SEQ ID NOs: 1422-1424, 1426, 1428, and 1429.

  • [0019]

    In certain preferred embodiments, the polypeptides and/or polynucleotides of the present invention are immunogenic, i.e., they are capable of eliciting an immune response, particularly a humoral and/or cellular immune response, as further described herein.

  • [0020]

    The present invention further provides fragments, variants and/or derivatives of the disclosed polypeptide and/or polynucleotide sequences, wherein the fragments, variants and/or derivatives preferably have a level of immunogenic activity of at least about 50%, preferably at least about 70% and more preferably at least about 90% of the level of immunogenic activity of a polypeptide sequence set forth in SEQ ID NOs: 1422-1424, 1426, 1428, and 1429 or a polypeptide sequence encoded by a polynucleotide sequence set forth in SEQ ID NOs: 1-1421, 1425, 1427, and 1430-3417.

  • [0021]

    The present invention further provides polynucleotides that encode a polypeptide described above, expression vectors comprising such polynucleotides and host cells transformed or transfected with such expression vectors.

  • [0022]

    Within other aspects, the present invention provides pharmaceutical compositions comprising a polypeptide or polynucleotide as described above and a physiologically acceptable carrier.

  • [0023]

    Within a related aspect of the present invention, the pharmaceutical compositions, e.g., vaccine compositions, are provided for prophylactic or therapeutic applications. Such compositions generally comprise an immunogenic polypeptide or polynucleotide of the invention and an immunostimulant, such as an adjuvant.

  • [0024]

    The present invention further provides pharmaceutical compositions that comprise: (a) an antibody or antigen-binding fragment thereof that specifically binds to a polypeptide of the present invention, or a fragment thereof; and (b) a physiologically acceptable carrier.

  • [0025]

    Within further aspects, the present invention provides pharmaceutical compositions comprising: (a) an antigen presenting cell that expresses a polypeptide as described above and (b) a pharmaceutically acceptable carrier or excipient. Illustrative antigen presenting cells include dendritic cells, macrophages, monocytes, fibroblasts and B cells.

  • [0026]

    Within related aspects, pharmaceutical compositions are provided that comprise: (a) an antigen presenting cell that expresses a polypeptide as described above and (b) an immunostimulant.

  • [0027]

    The present invention further provides, in other aspects, fusion proteins that comprise at least one polypeptide as described above, as well as polynucleotides encoding such fusion proteins, typically in the form of pharmaceutical compositions, e.g., vaccine compositions, comprising a physiologically acceptable carrier and/or an immunostimulant. The fusions proteins may comprise multiple immunogenic polypeptides or portions/variants thereof, as described herein, and may further comprise one or more polypeptide segments for facilitating the expression, purification and/or immunogenicity of the polypeptide(s).

  • [0028]

    Within further aspects, the present invention provides methods for stimulating an immune response in a patient, preferably a T cell response in a human patient, comprising administering a pharmaceutical composition described herein. The patient may be afflicted with colon cancer, in which case the methods provide treatment for the disease, or patient considered at risk for such a disease may be treated prophylactically.

  • [0029]

    Within further aspects, the present invention provides methods for inhibiting the development of a cancer in a patient, comprising administering to a patient a pharmaceutical composition as recited above. The patient may be afflicted with colon cancer, in which case the methods provide treatment for the disease, or patient considered at risk for such a disease may be treated prophylactically.

  • [0030]

    The present invention further provides, within other aspects, methods for removing tumor cells from a biological sample, comprising contacting a biological sample with T cells that specifically react with a polypeptide of the present invention, wherein the step of contacting is performed under conditions and for a time sufficient to permit the removal of cells expressing the protein from the sample.

  • [0031]

    Within related aspects, methods are provided for inhibiting the development of a cancer in a patient, comprising administering to a patient a biological sample treated as described above.

  • [0032]

    Methods are further provided, within other aspects, for stimulating and/or expanding T cells specific for a polypeptide of the present invention, comprising contacting T cells with one or more of: (i) a polypeptide as described above; (ii) a polynucleotide encoding such a polypeptide; and/or (iii) an antigen presenting cell that expresses such a polypeptide; under conditions and for a time sufficient to permit the stimulation and/or expansion of T cells. Isolated T cell populations comprising T cells prepared as described above are also provided.

  • [0033]

    Within further aspects, the present invention provides methods for inhibiting the development of a cancer in a patient, comprising administering to a patient an effective amount of a T cell population as described above.

  • [0034]

    The present invention further provides methods for inhibiting the development of a cancer in a patient, comprising the steps of: (a) incubating CD4+ and/or CD8+ T cells isolated from a patient with one or more of: (i) a polypeptide comprising at least an immunogenic portion of polypeptide disclosed herein; (ii) a polynucleotide encoding such a polypeptide; and (iii) an antigen-presenting cell that expressed such a polypeptide; and (b) administering to the patient an effective amount of the proliferated T cells, and thereby inhibiting the development of a cancer in the patient. Proliferated cells may, but need not, be cloned prior to administration to the patient.

  • [0035]

    Within further aspects, the present invention provides methods for determining the presence or absence of a cancer, preferably a colon cancer, in a patient comprising: (a) contacting a biological sample obtained from a patient with a binding agent that binds to a polypeptide as recited above; (b) detecting in the sample an amount of polypeptide that binds to the binding agent; and (c) comparing the amount of polypeptide with a predetermined cut-off value, and therefrom determining the presence or absence of a cancer in the patient. Within preferred embodiments, the binding agent is an antibody, more preferably a monoclonal antibody.

  • [0036]

    The present invention also provides, within other aspects, methods for monitoring the progression of a cancer in a patient. Such methods comprise the steps of: (a) contacting a biological sample obtained from a patient at a first point in time with a binding agent that binds to a polypeptide as recited above; (b) detecting in the sample an amount of polypeptide that binds to the binding agent; (c) repeating steps (a) and (b) using a biological sample obtained from the patient at a subsequent point in time; and (d) comparing the amount of polypeptide detected in step (c) with the amount detected in step (b) and therefrom monitoring the progression of the cancer in the patient.

  • [0037]

    The present invention further provides, within other aspects, methods for determining the presence or absence of a cancer in a patient, comprising the steps of: (a) contacting a biological sample, e.g., tumor sample, serum sample, etc., obtained from a patient with an oligonucleotide that hybridizes to a polynucleotide that encodes a polypeptide of the present invention; (b) detecting in the sample a level of a polynucleotide, preferably mRNA, that hybridizes to the oligonucleotide; and (c) comparing the level of polynucleotide that hybridizes to the oligonucleotide with a predetermined cut-off value, and therefrom determining the presence or absence of a cancer in the patient. Within certain embodiments, the amount of mRNA is detected via polymerase chain reaction using, for example, at least one oligonucleotide primer that hybridizes to a polynucleotide encoding a polypeptide as recited above, or a complement of such a polynucleotide. Within other embodiments, the amount of mRNA is detected using a hybridization technique, employing an oligonucleotide probe that hybridizes to a polynucleotide that encodes a polypeptide as recited above, or a complement of such a polynucleotide.

  • [0038]

    In related aspects, methods are provided for monitoring the progression of a cancer in a patient, comprising the steps of: (a) contacting a biological sample obtained from a patient with an oligonucleotide that hybridizes to a polynucleotide that encodes a polypeptide of the present invention; (b) detecting in the sample an amount of a polynucleotide that hybridizes to the oligonucleotide; (c) repeating steps (a) and (b) using a biological sample obtained from the patient at a subsequent point in time; and (d) comparing the amount of polynucleotide detected in step (c) with the amount detected in step (b) and therefrom monitoring the progression of the cancer in the patient.

  • [0039]

    Within further aspects, the present invention provides antibodies, such as monoclonal antibodies, that bind to a polypeptide as described above, as well as diagnostic kits comprising such antibodies. Diagnostic kits comprising one or more oligonucleotide probes or primers as described above are also provided.

  • [0040]

    These and other aspects of the present invention will become apparent upon reference to the following detailed description. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.

  • [0041]

    SEQ ID NO: 1-254 are the determined cDNA sequences described in Tables 2-10.

  • [0042]

    SEQ ID NO: 255 is the determined cDNA sequence for clone 63716879.

  • [0043]

    SEQ ID NO: 256 is the determined cDNA sequence for clone 63716880.

  • [0044]

    SEQ ID NO: 257 is the determined cDNA sequence for clone 63716882.

  • [0045]

    SEQ ID NO: 258 is the determined cDNA sequence for clone 63716883.

  • [0046]

    SEQ ID NO: 259 is the determined cDNA sequence for clone 63716884.

  • [0047]

    SEQ ID NO: 260 is the determined cDNA sequence for clone 63716885.

  • [0048]

    SEQ ID NO: 261 is the determined cDNA sequence for clone 63716886.

  • [0049]

    SEQ ID NO: 262 is the determined cDNA sequence for clone 63716887.

  • [0050]

    SEQ ID NO: 263 is the determined cDNA sequence for clone 63716888.

  • [0051]

    SEQ ID NO: 264 is the determined cDNA sequence for clone 63716889.

  • [0052]

    SEQ ID NO: 265 is the determined cDNA sequence for clone 63716890.

  • [0053]

    SEQ ID NO: 266 is the determined cDNA sequence for clone 63716891.

  • [0054]

    SEQ ID NO: 267 is the determined cDNA sequence for clone 63716892.

  • [0055]

    SEQ ID NO: 268 is the determined cDNA sequence for clone 63716894.

  • [0056]

    SEQ ID NO: 269 is the determined cDNA sequence for clone 63716895.

  • [0057]

    SEQ ID NO: 270 is the determined cDNA sequence for clone 63716896.

  • [0058]

    SEQ ID NO: 271 is the determined cDNA sequence for clone 63716897.

  • [0059]

    SEQ ID NO: 272 is the determined cDNA sequence for clone 63716898.

  • [0060]

    SEQ ID NO: 273 is the determined cDNA sequence for clone 63716899.

  • [0061]

    SEQ ID NO: 274 is the determined cDNA sequence for clone 63716901.

  • [0062]

    SEQ ID NO: 275 is the determined cDNA sequence for clone 63716902.

  • [0063]

    SEQ ID NO: 276 is the determined cDNA sequence for clone 63716903.

  • [0064]

    SEQ ID NO: 277 is the determined cDNA sequence for clone 63716904.

  • [0065]

    SEQ ID NO: 278 is the determined cDNA sequence for clone 63716905.

  • [0066]

    SEQ ID NO: 279 is the determined cDNA sequence for clone 63716906.

  • [0067]

    SEQ ID NO: 280 is the determined cDNA sequence for clone 63716907.

  • [0068]

    SEQ ID NO: 281 is the determined cDNA sequence for clone 63716908.

  • [0069]

    SEQ ID NO: 282 is the determined cDNA sequence for clone 63716909.

  • [0070]

    SEQ ID NO: 283 is the determined cDNA sequence for clone 63716910.

  • [0071]

    SEQ ID NO: 284 is the determined cDNA sequence for clone 63716911.

  • [0072]

    SEQ ID NO: 285 is the determined cDNA sequence for clone 63716912.

  • [0073]

    SEQ ID NO: 286 is the determined cDNA sequence for clone 63716914.

  • [0074]

    SEQ ID NO: 287 is the determined cDNA sequence for clone 63716915.

  • [0075]

    SEQ ID NO: 288 is the determined cDNA sequence for clone 63716916.

  • [0076]

    SEQ ID NO: 289 is the determined cDNA sequence for clone 63716918.

  • [0077]

    SEQ ID NO: 290 is the determined cDNA sequence for clone 63716919.

  • [0078]

    SEQ ID NO: 291 is the determined cDNA sequence for clone 63716920.

  • [0079]

    SEQ ID NO: 292 is the determined cDNA sequence for clone 63716922.

  • [0080]

    SEQ ID NO: 293 is the determined cDNA sequence for clone 63716923.

  • [0081]

    SEQ ID NO: 294 is the determined cDNA sequence for clone 63716924.

  • [0082]

    SEQ ID NO: 295 is the determined cDNA sequence for clone 63716925.

  • [0083]

    SEQ ID NO: 296 is the determined cDNA sequence for clone 63716926.

  • [0084]

    SEQ ID NO: 297 is the determined cDNA sequence for clone 63716927.

  • [0085]

    SEQ ID NO: 298 is the determined cDNA sequence for clone 63716928.

  • [0086]

    SEQ ID NO: 299 is the determined cDNA sequence for clone 63716929.

  • [0087]

    SEQ ID NO: 300 is the determined cDNA sequence for clone 63716930.

  • [0088]

    SEQ ID NO: 301 is the determined cDNA sequence for clone 63716931.

  • [0089]

    SEQ ID NO: 302 is the determined cDNA sequence for clone 63716932.

  • [0090]

    SEQ ID NO: 303 is the determined cDNA sequence for clone 63716933.

  • [0091]

    SEQ ID NO: 304 is the determined cDNA sequence for clone 63716934.

  • [0092]

    SEQ ID NO: 305 is the determined cDNA sequence for clone 63716935.

  • [0093]

    SEQ ID NO: 306 is the determined cDNA sequence for clone 63716936.

  • [0094]

    SEQ ID NO: 307 is the determined cDNA sequence for clone 63716937.

  • [0095]

    SEQ ID NO: 308 is the determined cDNA sequence for clone 63716938.

  • [0096]

    SEQ ID NO: 309 is the determined cDNA sequence for clone 63716939.

  • [0097]

    SEQ ID NO: 310 is the determined cDNA sequence for clone 63716940.

  • [0098]

    SEQ ID NO: 311 is the determined cDNA sequence for clone 63716941.

  • [0099]

    SEQ ID NO: 312 is the determined cDNA sequence for clone 63716942.

  • [0100]

    SEQ ID NO: 313 is the determined cDNA sequence for clone 63716943.

  • [0101]

    SEQ ID NO: 314 is the determined cDNA sequence for clone 63716944.

  • [0102]

    SEQ ID NO: 315 is the determined cDNA sequence for clone 63716945.

  • [0103]

    SEQ ID NO: 316 is the determined cDNA sequence for clone 63716946.

  • [0104]

    SEQ ID NO: 317 is the determined cDNA sequence for clone 63716948.

  • [0105]

    SEQ ID NO: 318 is the determined cDNA sequence for clone 63716949.

  • [0106]

    SEQ ID NO: 319 is the determined cDNA sequence for clone 63716950.

  • [0107]

    SEQ ID NO: 320 is the determined cDNA sequence for clone 63716951.

  • [0108]

    SEQ ID NO: 321 is the determined cDNA sequence for clone 63716953.

  • [0109]

    SEQ ID NO: 322 is the determined cDNA sequence for clone 63716954.

  • [0110]

    SEQ ID NO: 323 is the determined cDNA sequence for clone 63716955.

  • [0111]

    SEQ ID NO: 324 is the determined cDNA sequence for clone 63716956.

  • [0112]

    SEQ ID NO: 325 is the determined cDNA sequence for clone 63716957.

  • [0113]

    SEQ ID NO: 326 is the determined cDNA sequence for clone 63716958.

  • [0114]

    SEQ ID NO: 327 is the determined cDNA sequence for clone 63716959.

  • [0115]

    SEQ ID NO: 328 is the determined cDNA sequence for clone 63716960.

  • [0116]

    SEQ ID NO: 329 is the determined cDNA sequence for clone 63716961.

  • [0117]

    SEQ ID NO: 330 is the determined cDNA sequence for clone 63716962.

  • [0118]

    SEQ ID NO: 331 is the determined cDNA sequence for clone 63716963.

  • [0119]

    SEQ ID NO: 332 is the determined cDNA sequence for clone 63716964.

  • [0120]

    SEQ ID NO: 333 is the determined cDNA sequence for clone 63716965.

  • [0121]

    SEQ ID NO: 334 is the determined cDNA sequence for clone 63716966.

  • [0122]

    SEQ ID NO: 335 is the determined cDNA sequence for clone 63716967.

  • [0123]

    SEQ ID NO: 336 is the determined cDNA sequence for clone 63716968.

  • [0124]

    SEQ ID NO: 337 is the determined cDNA sequence for clone 63716970.

  • [0125]

    SEQ ID NO: 338 is the determined cDNA sequence for clone 63716971.

  • [0126]

    SEQ ID NO: 339 is the determined cDNA sequence for clone 63716786.

  • [0127]

    SEQ ID NO: 340 is the determined cDNA sequence for clone 63716787.

  • [0128]

    SEQ ID NO: 341 is the determined cDNA sequence for clone 63716788.

  • [0129]

    SEQ ID NO: 342 is the determined cDNA sequence for clone 63716789.

  • [0130]

    SEQ ID NO: 343 is the determined cDNA sequence for clone 63716790.

  • [0131]

    SEQ ID NO: 344 is the determined cDNA sequence for clone 63716792.

  • [0132]

    SEQ ID NO: 345 is the determined cDNA sequence for clone 63716793.

  • [0133]

    SEQ ID NO: 346 is the determined cDNA sequence for clone 63716794.

  • [0134]

    SEQ ID NO: 347 is the determined cDNA sequence for clone 63716798.

  • [0135]

    SEQ ID NO: 348 is the determined cDNA sequence for clone 63716799.

  • [0136]

    SEQ ID NO: 349 is the determined cDNA sequence for clone 63716802.

  • [0137]

    SEQ ID NO: 350 is the determined cDNA sequence for clone 63716803.

  • [0138]

    SEQ ID NO: 351 is the determined cDNA sequence for clone 63716806.

  • [0139]

    SEQ ID NO: 352 is the determined cDNA sequence for clone 63716807.

  • [0140]

    SEQ ID NO: 353 is the determined cDNA sequence for clone 63716810.

  • [0141]

    SEQ ID NO: 354 is the determined cDNA sequence for clone 63716811.

  • [0142]

    SEQ ID NO: 355 is the determined cDNA sequence for clone 63716812.

  • [0143]

    SEQ ID NO: 356 is the determined cDNA sequence for clone 63716813.

  • [0144]

    SEQ ID NO: 357 is the determined cDNA sequence for clone 63716814.

  • [0145]

    SEQ ID NO: 358 is the determined cDNA sequence for clone 63716815.

  • [0146]

    SEQ ID NO: 359 is the determined cDNA sequence for clone 63716816.

  • [0147]

    SEQ ID NO: 360 is the determined cDNA sequence for clone 63716817.

  • [0148]

    SEQ ID NO: 361 is the determined cDNA sequence for clone 63716818.

  • [0149]

    SEQ ID NO: 362 is the determined cDNA sequence for clone 63716819.

  • [0150]

    SEQ ID NO: 363 is the determined cDNA sequence for clone 63716820.

  • [0151]

    SEQ ID NO: 364 is the determined cDNA sequence for clone 63716822.

  • [0152]

    SEQ ID NO: 365 is the determined cDNA sequence for clone 63716824.

  • [0153]

    SEQ ID NO: 366 is the determined cDNA sequence for clone 63716825.

  • [0154]

    SEQ ID NO: 367 is the determined cDNA sequence for clone 63716826.

  • [0155]

    SEQ ID NO: 368 is the determined cDNA sequence for clone 63716828.

  • [0156]

    SEQ ID NO: 369 is the determined cDNA sequence for clone 63716829.

  • [0157]

    SEQ ID NO: 370 is the determined cDNA sequence for clone 63716830.

  • [0158]

    SEQ ID NO: 371 is the determined cDNA sequence for clone 63716831.

  • [0159]

    SEQ ID NO: 372 is the determined cDNA sequence for clone 63716834.

  • [0160]

    SEQ ID NO: 373 is the determined cDNA sequence for clone 63716835.

  • [0161]

    SEQ ID NO: 374 is the determined cDNA sequence for clone 63716836.

  • [0162]

    SEQ ID NO: 375 is the determined cDNA sequence for clone 63716837.

  • [0163]

    SEQ ID NO: 376 is the determined cDNA sequence for clone 63716838.

  • [0164]

    SEQ ID NO: 377 is the determined cDNA sequence for clone 63716839.

  • [0165]

    SEQ ID NO: 378 is the determined cDNA sequence for clone 63716842.

  • [0166]

    SEQ ID NO: 379 is the determined cDNA sequence for clone 63716843.

  • [0167]

    SEQ ID NO: 380 is the determined cDNA sequence for clone 63716844.

  • [0168]

    SEQ ID NO: 381 is the determined cDNA sequence for clone 63716846.

  • [0169]

    SEQ ID NO: 382 is the determined cDNA sequence for clone 63716847.

  • [0170]

    SEQ ID NO: 383 is the determined cDNA sequence for clone 63716848.

  • [0171]

    SEQ ID NO: 384 is the determined cDNA sequence for clone 63716851.

  • [0172]

    SEQ ID NO: 385 is the determined cDNA sequence for clone 63716852.

  • [0173]

    SEQ ID NO: 386 is the determined cDNA sequence for clone 63716853.

  • [0174]

    SEQ ID NO: 387 is the determined cDNA sequence for clone 63716855.

  • [0175]

    SEQ ID NO: 388 is the determined cDNA sequence for clone 63716858.

  • [0176]

    SEQ ID NO: 389 is the determined cDNA sequence for clone 63716860.

  • [0177]

    SEQ ID NO: 390 is the determined cDNA sequence for clone 63716861.

  • [0178]

    SEQ ID NO: 391 is the determined cDNA sequence for clone 63716862.

  • [0179]

    SEQ ID NO: 392 is the determined cDNA sequence for clone 63716863.

  • [0180]

    SEQ ID NO: 393 is the determined cDNA sequence for clone 63716865.

  • [0181]

    SEQ ID NO: 394 is the determined cDNA sequence for clone 63716866.

  • [0182]

    SEQ ID NO: 395 is the determined cDNA sequence for clone 63716868.

  • [0183]

    SEQ ID NO: 396 is the determined cDNA sequence for clone 63716869.

  • [0184]

    SEQ ID NO: 397 is the determined cDNA sequence for clone 63716870.

  • [0185]

    SEQ ID NO: 398 is the determined cDNA sequence for clone 63716871.

  • [0186]

    SEQ ID NO: 399 is the determined cDNA sequence for clone 63716873.

  • [0187]

    SEQ ID NO: 400 is the determined cDNA sequence for clone 63716875.

  • [0188]

    SEQ ID NO: 401 is the determined cDNA sequence for clone 63716876.

  • [0189]

    SEQ ID NO: 402 is the determined cDNA sequence for clone 63716877.

  • [0190]

    SEQ ID NO: 403 is the determined cDNA sequence for clone 63716878.

  • [0191]

    SEQ ID NO: 404 is the determined cDNA sequence for clone 63717158.

  • [0192]

    SEQ ID NO: 405 is the determined cDNA sequence for clone 63717160.

  • [0193]

    SEQ ID NO: 406 is the determined cDNA sequence for clone 63717161.

  • [0194]

    SEQ ID NO: 407 is the determined cDNA sequence for clone 63717163.

  • [0195]

    SEQ ID NO: 408 is the determined cDNA sequence for clone 63717164.

  • [0196]

    SEQ ID NO: 409 is the determined cDNA sequence for clone 63717165.

  • [0197]

    SEQ ID NO: 410 is the determined cDNA sequence for clone 63717166.

  • [0198]

    SEQ ID NO: 411 is the determined cDNA sequence for clone 63717167.

  • [0199]

    SEQ ID NO: 412 is the determined cDNA sequence for clone 63717169.

  • [0200]

    SEQ ID NO: 413 is the determined cDNA sequence for clone 63717171.

  • [0201]

    SEQ ID NO: 414 is the determined cDNA sequence for clone 63717172.

  • [0202]

    SEQ ID NO: 415 is the determined cDNA sequence for clone 63717173.

  • [0203]

    SEQ ID NO: 416 is the determined cDNA sequence for clone 63717174.

  • [0204]

    SEQ ID NO: 417 is the determined cDNA sequence for clone 63717175.

  • [0205]

    SEQ ID NO: 418 is the determined cDNA sequence for clone 63717176.

  • [0206]

    SEQ ID NO: 419 is the determined cDNA sequence for clone 63717177.

  • [0207]

    SEQ ID NO: 420 is the determined cDNA sequence for clone 63717178.

  • [0208]

    SEQ ID NO: 421 is the determined cDNA sequence for clone 63717179.

  • [0209]

    SEQ ID NO: 422 is the determined cDNA sequence for clone 63717180.

  • [0210]

    SEQ ID NO: 423 is the determined cDNA sequence for clone 63717181.

  • [0211]

    SEQ ID NO: 424 is the determined cDNA sequence for clone 63717182.

  • [0212]

    SEQ ID NO: 425 is the determined cDNA sequence for clone 63717183.

  • [0213]

    SEQ ID NO: 426 is the determined cDNA sequence for clone 63717184.

  • [0214]

    SEQ ID NO: 427 is the determined cDNA sequence for clone 63717186.

  • [0215]

    SEQ ID NO: 428 is the determined cDNA sequence for clone 63717187.

  • [0216]

    SEQ ID NO: 429 is the determined cDNA sequence for clone 63717189.

  • [0217]

    SEQ ID NO: 430 is the determined cDNA sequence for clone 63717190.

  • [0218]

    SEQ ID NO: 431 is the determined cDNA sequence for clone 63717191.

  • [0219]

    SEQ ID NO: 432 is the determined cDNA sequence for clone 63717192.

  • [0220]

    SEQ ID NO: 433 is the determined cDNA sequence for clone 63717193.

  • [0221]

    SEQ ID NO: 434 is the determined cDNA sequence for clone 63717194.

  • [0222]

    SEQ ID NO: 435 is the determined cDNA sequence for clone 63717197.

  • [0223]

    SEQ ID NO: 436 is the determined cDNA sequence for clone 63717199.

  • [0224]

    SEQ ID NO: 437 is the determined cDNA sequence for clone 63717200.

  • [0225]

    SEQ ID NO: 438 is the determined cDNA sequence for clone 63717201.

  • [0226]

    SEQ ID NO: 439 is the determined cDNA sequence for clone 63717202.

  • [0227]

    SEQ ID NO: 440 is the determined cDNA sequence for clone 63717203.

  • [0228]

    SEQ ID NO: 441 is the determined cDNA sequence for clone 63717204.

  • [0229]

    SEQ ID NO: 442 is the determined cDNA sequence for clone 63717205.

  • [0230]

    SEQ ID NO: 443 is the determined cDNA sequence for clone 63717206.

  • [0231]

    SEQ ID NO: 444 is the determined cDNA sequence for clone 63717207.

  • [0232]

    SEQ ID NO: 445 is the determined cDNA sequence for clone 63717208.

  • [0233]

    SEQ ID NO: 446 is the determined cDNA sequence for clone 63717209.

  • [0234]

    SEQ ID NO: 447 is the determined cDNA sequence for clone 63717211.

  • [0235]

    SEQ ID NO: 448 is the determined cDNA sequence for clone 63717212.

  • [0236]

    SEQ ID NO: 449 is the determined cDNA sequence for clone 63717213.

  • [0237]

    SEQ ID NO: 450 is the determined cDNA sequence for clone 63717214.

  • [0238]

    SEQ ID NO: 451 is the determined cDNA sequence for clone 63717215.

  • [0239]

    SEQ ID NO: 452 is the determined cDNA sequence for clone 63717216.

  • [0240]

    SEQ ID NO: 453 is the determined cDNA sequence for clone 63717217.

  • [0241]

    SEQ ID NO: 454 is the determined cDNA sequence for clone 63717218.

  • [0242]

    SEQ ID NO: 455 is the determined cDNA sequence for clone 63717219.

  • [0243]

    SEQ ID NO: 456 is the determined cDNA sequence for clone 63717221.

  • [0244]

    SEQ ID NO: 457 is the determined cDNA sequence for clone 63717222.

  • [0245]

    SEQ ID NO: 458 is the determined cDNA sequence for clone 63717223.

  • [0246]

    SEQ ID NO: 459 is the determined cDNA sequence for clone 63717224.

  • [0247]

    SEQ ID NO: 460 is the determined cDNA sequence for clone 63717227.

  • [0248]

    SEQ ID NO: 461 is the determined cDNA sequence for clone 63717228.

  • [0249]

    SEQ ID NO: 462 is the determined cDNA sequence for clone 63717229.

  • [0250]

    SEQ ID NO: 463 is the determined cDNA sequence for clone 63717231.

  • [0251]

    SEQ ID NO: 464 is the determined cDNA sequence for clone 63717233.

  • [0252]

    SEQ ID NO: 465 is the determined cDNA sequence for clone 63717234.

  • [0253]

    SEQ ID NO: 466 is the determined cDNA sequence for clone 63717235.

  • [0254]

    SEQ ID NO: 467 is the determined cDNA sequence for clone 63717236.

  • [0255]

    SEQ ID NO: 468 is the determined cDNA sequence for clone 63717237.

  • [0256]

    SEQ ID NO: 469 is the determined cDNA sequence for clone 63717238.

  • [0257]

    SEQ ID NO: 470 is the determined cDNA sequence for clone 63717239.

  • [0258]

    SEQ ID NO: 471 is the determined cDNA sequence for clone 63717240.

  • [0259]

    SEQ ID NO: 472 is the determined cDNA sequence for clone 63717241.

  • [0260]

    SEQ ID NO: 473 is the determined cDNA sequence for clone 63717242.

  • [0261]

    SEQ ID NO: 474 is the determined cDNA sequence for clone 63717243.

  • [0262]

    SEQ ID NO: 475 is the determined cDNA sequence for clone 63717244.

  • [0263]

    SEQ ID NO: 476 is the determined cDNA sequence for clone 63717246.

  • [0264]

    SEQ ID NO: 477 is the determined cDNA sequence for clone 63717248.

  • [0265]

    SEQ ID NO: 478 is the determined cDNA sequence for clone 63717250.

  • [0266]

    SEQ ID NO: 479 is the determined cDNA sequence for clone 63716972.

  • [0267]

    SEQ ID NO: 480 is the determined cDNA sequence for clone 63716973.

  • [0268]

    SEQ ID NO: 481 is the determined cDNA sequence for clone 63716975.

  • [0269]

    SEQ ID NO: 482 is the determined cDNA sequence for clone 63716976.

  • [0270]

    SEQ ID NO: 483 is the determined cDNA sequence for clone 63716977.

  • [0271]

    SEQ ID NO: 484 is the determined cDNA sequence for clone 63716978.

  • [0272]

    SEQ ID NO: 485 is the determined cDNA sequence for clone 63716979.

  • [0273]

    SEQ ID NO: 486 is the determined cDNA sequence for clone 63716980.

  • [0274]

    SEQ ID NO: 487 is the determined cDNA sequence for clone 63716981.

  • [0275]

    SEQ ID NO: 488 is the determined cDNA sequence for clone 63716982.

  • [0276]

    SEQ ID NO: 489 is the determined cDNA sequence for clone 63716984.

  • [0277]

    SEQ ID NO: 490 is the determined cDNA sequence for clone 63716985.

  • [0278]

    SEQ ID NO: 491 is the determined cDNA sequence for clone 63716986.

  • [0279]

    SEQ ID NO: 492 is the determined cDNA sequence for clone 63716987.

  • [0280]

    SEQ ID NO: 493 is the determined cDNA sequence for clone 63716988.

  • [0281]

    SEQ ID NO: 494 is the determined cDNA sequence for clone 63716989.

  • Я люблю тебя, - шептал коммандер.  - Я любил тебя. У нее свело желудок.

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