USO0RE36713E

United States Patent [19]

[11] E

Vogelstein et al.

[45] Reissued Date of Patent:

[54]

APC GENE AND NUCLEIC ACID PROBES

Patent Number:

Re. 36,713 May 23, 2000

[52]

US. Cl. ................ .. 536/23.5; 536/2431; 536/2433;

[58]

Field of Search .............................. .. 536/23.5, 24.31,

DERIVED THEREFROM

536/231; 435/6; 935/6; 935/8; 935/9

[75] Inventors: Bert Vogelstein, Baltimore; Kenneth W. Kinzler, BelAir, both of Md.; Hans Albertson, Salt Lake City, Utah; Rakesh Anand, Sandbach, United

536/231, 24.33; 935/6, 8, 9; 435/6 [56]

References Cited

Kingdom; Mary Carlson; Joanna

U.S. PATENT DOCUMENTS

Groden, both of Salt Lake City, Utah; 5,098,823 5,137,806

Philip John Hedge, Knutsford, United Kingdom; Geo?' Joslyn, Salt Lake City, Utah; Alexander Fred Markham, CreWe, United Kingdom; Yusuke

FOREIGN PATENT DOCUMENTS 89/01481

Nakamura, Tokyo, Japan; Andrew Thliveris; Raymond White, both of

Baltimore, Md.; The University of Utah, Salt Lake City, Utah; Zeneca, United Kingdom; Cancer Institute,

Tokyo, Japan [21] Appl. No.: 08/726,870 [22]

Filed:

2/1989

WIPO.

OTHER PUBLICATIONS

Salt Lake City, Utah

[73] Assignees: The Johns Hopkins University,

3/1992 Bodmer et al. ........................... .. 435/6 8/1992 LeMaistre et al. ....................... .. 436/6

Groden, et al., Cell, vol. 66, pp. 589—600, 1991. Joslyn, et al., Cell, vol. 66, pp. 601—613, 1991. KinZler, et al., Science, vol. 253, pp. 661—665, 1991. Nishisho et al., Science, vol. 253, pp. 665—669, 1991. Orita, et al., Genomics, vol. 5, pp. 874—879, 1989. Stanbridge, et al., Science, vol. 247, pp. 12—13, 1990. Bodmer et al., 1987 Nature 328:614. Fearen et al 1990 Science 247:49. Baker et al. 1989 Science 244:217.

Oct. 4, 1996 Related US. Patent Documents

Reissue of:

Primary Examiner—David L. Fitzgerald

[64]

[30]

Patent No.:

5,352,775

Attorney, Agent, or Firm—Banner & Witcoff, Ltd.

Issued:

Oct. 4, 1994

Appl. No.:

07/741,940

[57]

Filed:

Aug. 8, 1991

Foreign Application Priority Data

Jan. 16, 1991

[GB]

United Kingdom ................. .. 9100962

Jan. 16, 1991 Jan. 16, 1991

[GB] [GB]

United Kingdom United Kingdom

Jan. 16, 1991

[GB]

United Kingdom ................. .. 9100975

[51]

9100963 9100974

ABSTRACT

A human gene termed APC is disclosed. Methods and kits are provided for assessing mutations of the APC gene in hum tissues and body samples. APC mutations are found in familial adenomatous polyposis patients as Wel as in spo radic colorectal cancer patents. APC is expressed in most normal tissue. These results suggest that APC is a tumor suppressor.

Int. Cl.7 ...................... .. C12N 15/12; C07K 14/435;

C12Q 01/68

10 Claims, 40 Drawing Sheets

U.S. Patent

May 23, 2000

Sheet 2 0f 40

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U.S. Patent

May 23, 2000

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Sheet 5 0f 40

Re. 36,713

U.S. Patent

May 23, 2000

Re. 36,713

Sheet 7 0f 40

Amended Figure 3B PLMFSRCTSV SSLDSFESRS IASSVQSEPC SGMVSGIISP SDLPDSPGQT MPPSRSKTPP 1450 1460 1470 1480 1490 1500 PPPQTAQTKR EVPKNKAPTA EKRESGPKQA AVNAAVQRVQ VLPDADTLLH FATESTPDGF 1510 1520 1530 1540 1550 1560 SCSSSLSALS LDEPFIQKDV ELRIMPPVQE NDNGNETESE QPKESNENQE KEAEKTIDSE 1570 1580 1590 1600 1610 1620 KDLLDDSDDD DIEILEECII SAMPTKSSRK AKKPAQTASK LPPPVARKPS QLPVYKLLPS 1630 1640 1650 1660 1670 1680 QNRLQPQKHV SFTPGDDMPR VYCVEGTPIN FSTATSLSDL TIESPPNELA AGEGVRGGAQ 1690 1700 1710 1720 1730 1740 SGEFEKRDTI PTEGRSTDEA QGGKTSSVTI PELDDNKAEE GDILAECINS AMPKGKSHKP 1750 1760 1770 1780 1790 1800 FRVKKIMDQV QQASASSSAP NKNQLDGKKK KPTSPVKPIP QNTEYRTRVR KNADSKNNLN 1810 1820 1830 1840 1850 1860 AERVFSDNKD SKKQNLKNNS KDFNDKLPNN EDRVRGSFAF DSPHHYTPIE GTPYCFSRND 1870 1880 1890 1900 1910 193.0 SLSSLDFDDD DVDLSREKAE LRKAKENKES EAKVTSHTEL TSNQQSANKT QAIAKQPINR 4 1930 1940 1950 1960 1970 1980 GQPKPILQKQ STFPQSSKDI PDRGAATDEK LQNFAIEN‘I'P VCFSHNSSLS SLSDIDQENN 1990 2000 2010 2020 2030 2040 NKENEPIKET EPPDSQGEPS KPQASGYAPK SFHVEDTPVC FSRNSSLSSL SIDSEDDLLQ 2050 2060 2070 2080 2090 2100 ECISSAMPKK KKPSRLKGDN EKHSPRNMGG ILGEDLTLDL KDIQRPDSEH GLSPDSENFD 2110 2120 2130 2140 2150 2160 WKAIQEGANS IVSSLHQAAA AACLSRQASS DSDSILSLKS GISLGSPFHL TPDQEEKPFT 2170 2180 2190 2200 2210 2220 SNKGPRILKP GEKSTLETKK IESESKGIKG GKKVYKSLIT GKVRSNSEIS GQMKQPLQAN 2230 2240 2250 2260 2270 2280 MPSISRGRTM IHIPGVRNSS SSTSPVSKKG PPLKTPASKS PSEGQTATTS PRGAKPSVKS 2290 2300 2310 2320 2330 2340 ELSPVARQTS QIGGSSKAPS RSGSRDSTPS RPAQQPLSRP IQSPGRNSIS PGRNGISPPN 2350 2360 2370 2380 2390 2400 KLSQLPRTSS PSTASTKSSG SGKMSYTSPG RQMSQQNLTK QTGLSKNASS IPRSESASKG 2410 2420 2430 2440 2450 2460 LNQMN'NGNGA NKKVELSRMS STKSSGSESD RSERPVLVRQ STFIKEAPSP TLRRKLEESA 2470 2480 2490 2500 2510 2520 SFESLSPSSR PASPTRSQAQ TPV'LSPSLPD MSLSTHSSVQ AGGWRKLPPN LSPTIEYNDG 2530 2540 2550 2560 2570 2580 RPAKRHDIAR SHSESPSRLP INRSGTWKRE HSKHSSSLPR VSTWRRTGSS SSILSASSES 2590 SEKAKSEDEK 2650

2600

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2630

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HVNSISGTKQ SKENQVSAKG TWRKIKENEF SPTNSTSQTV SSGATNGAES 2660

2670

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KTLIYQMAPA VSKTEDVW'VR IEDCPINNPR SGRSPTGNTP PVIDSVSEKA NPNIKDSKDN 2710

2720

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QAKQNVGNGS VPMR'I‘VGLEN RLNSFIQVDA PDQKGTEIKP GQNNPVPVSE TNESSIVERT 2770 2780 2790 2800 2810 2820 PFSSSSSSKH SSPSGTVAAR VTPFNYNPSP RKSSADSTSA RPSQIPTPVN NNTKKRDSKT

U.S. Patent

May 23, 2000

Sheet 8 0f 40

Re. 36,713

Amended Figure BC 2830 2840 DSTESSGTQS PKRHSGSYLV TSV*

U.S. Patent

May 23, 2000

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U.S. Patent

May 23, 2000

Sheet 12 0f 40

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U.S. Patent

May 23, 2000

Sheet 13 0f 40

Re. 36,713

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U.S. Patent

May 23, 2000

Sheet 14 0f 40

Re. 36,713

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U.S. Patent

May 23, 2000

Sheet 15 0f 40

Re. 36,713

Amended Figure 7A 23 ATGGCTGCAGCTTCATATGATCAGTTGTTAAAGCAAGTTGAGGCACTGAAGATGGAGAAC

l

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20

83 TCAAATCTTCGACAAGAGCTAGAAGATAATTCCAA'I‘CATCT'I‘ACAAAACTGGAAAC'I‘GAG

21

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263 AA'I‘TTCCCTGGAGTAAAAC‘I'GCGGTCAAAAATGTCCCTCCGTTCTTATGGAAGCCGGGAA

81

N

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202

60

203 GCTTCTTCTGGACAGATTGATTTATTAGAGCGTCTTAAAGAGCTTAACT'I‘AGATAGCAGT

61

142

40

143 GCATCTAA'I‘ATGAAGGAAGTAC‘I‘TAAACAACTACAAGGAAGTATTGAAGATGAAGCTATG

41

82

E

262

80

322

100

323 GGATCTG‘I‘ATCAAGCCGTTCTGGAGAGTGCAGTCCTGTTCCTATGGGTTCAT'I'TCCAAGA

382

101

120

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383 AGAGGGTTTGTAAATGGAAGCAGAGAAAGTACTGGATATTTAGAAGAACTTGAGAAAGAG

442

U.S. Patent

May 23, 2000

Sheet 16 0f 40

Re. 36,713

Amended F1'8"’6 7B 121.

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443 AGGTCATTGCTTCTTGCTGATCTTGACAAAGAAGAAAAGGAAAAAGACTGGTATTACGCT

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141

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503 CAACTTCAGAATCTCACTAAAAGAATAGATAGTCTTCCT‘I‘TAACTGAAAATTTTTCCTTA

562

161

180

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563 CAAACAGATATGACCAGAAGGCAATTGGAATATGAAGCAAGGCAAATCAGAGTTGCGATG

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623 GAAGAACAACTAGGTACCTGCCAGGATATGGAAAAACGAGCACAGCGAAGAATAGCCAGA

682

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683 ATTCAGCAAATCGAAAAGGACATAC'I'TCGTATACGACAGCTTTTACAGTCCCAAGCAACA

742

221

240

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743 GAAGCAGAGAGGTCATCTCAGAACAAGCATGAAACCGGCTCACATGATGCTGAGCGGCAG

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U.S. Patent

May 23, 2000

Sheet 17 0f 40

Re. 36,713

Amended Figure 7C 803 AATGAAGGTCAAGGAGTGGGAGAAATCAACATGGCAACTTCTGGTAATGGTCAGGGTTCA

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863 ACTACACGAATGGACCATGAAACAGCCAGTGTTTTGAGTTCTAGTAGCACACACTCTGCA

922

281T'1‘RMDHETASVLSSSSTHSA

300

923 CCTCGAAGGCTGACAAGTCATCTGGGAACCAAGgtggaaatggtgtattcattgttgtca

982

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320

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1043 caagacagctgtatatccatgcgacagtctggatgtcttcctctcctcatccagctttta

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1103 catggcaatgacaaagactctgtattgttgggagattcccggggcagtaaagaggctcgg

1162

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1163 gccagggccagtgcagcactc:acaacatcattcactcacagcctgatgacaagagaggc

380

1222

U.S. Patent

May 23, 2000

Sheet 18 0f 40

Re. 36,713

Amended Figure 7D 381

a

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1223 aggcgtgaaatccgagtccttcatcttttgga.acagATACGCGCTTACTGTGAAACCTGT

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1343 CCTGTTGAACATCAGATCTGTCCTGC'I‘GTGTG'I‘GTTCTAATGAAACTTTCATTTGATGAA

441

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460

1403 GAGCATAGACATGCAATGAATGAACTAGGGGGACTACAGGCCATTGCAGAATTATTGCAA

461

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1523 G-CTGGAATGGCTT'I‘GACAAACTTGAC'I‘TTTGGAGATGTAGCCAACAAGGCTACGCTATGC

501

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1462

480

1463 GTGGACTGTGAAATGTACGGGCTTACTAATGACCACTACAGTATTACACTAAGACGATAT

481

1402

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1522

500

1582

520

U.S. Patent

May 23, 2000

Sheet 19 0f 40

Re. 36,713

Amended Figure 7E 1 5 8 3 TCTAT GAAAGGCT GCAT GAGAGCACTTGTGGCCCAACTAAAAT CT GAAAGT GAAGACT'I‘A

521

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1 6 4 3 CAGCAGGTTATT GCAAGT GTTTT GAGGAATTT GT CTTGGCGAGCAGAT GTAAATAGTAAA

541

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1 '7 O 3 AAGACGTTGCGAGAAGTTGGAAGTGTGAAAGCATTGATGGAATGTGCTTTAGAAGTTAAA

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1883 ACTTACCGGAGCCAGACAAACACTTTAGCCATTATTGAAAGTGGAGGTGGGATATTACGG

1942

621TYRSQTNTLAIIESGGGILR

640