United States Patent
`Skolnick et al.
`
`[19]
`
`[54] 170-LINKED BREAST AND OVARIAN
`CANCER SUSCEPTIBILITY GENE
`
`[75]
`
`Inventors: Mark H. Skolnick; David E. Goldgar;
`Yoshio Mild; Jeff Swenson; Alexander
`Kamb; Keith D. Harslnnan; Donna
`M. Shattuck-Eidens; Sean V.
`Tavtigian. all of Salt Lake City, Utah;
`Roger W. Wiseman; P. Andrew
`Futreal, both of Durham. N.C.
`
`[73] Assignees: Myriad Genetics, Inc.; University of
`Utah Research Foundation, both of
`Salt Lake City. Utah; The United
`States of America as represented by
`the Department of Health and
`Human Services. Washington. D.C.
`
`[21] Appl. No.: 488,011
`
`[22] Filed:
`
`Jan. 5, 1!J!J6
`
`Related U.S. Application Data
`
`[51]
`
`[63] Continuation-in-part of Ser. No. 409,305, Mar. 24, 1995,
`abandoned, which is a continuation-in-part of Ser. No.
`348,824, Nov. 29, 1994, abandoned, which is a continuation(cid:173)
`in-part of Ser. No. 308,104, Sep. 16, 1994, abandoned,
`which is a continuation-in-part of Ser. No. 300,266, Sep. 2,
`1994, abandoned, which is a continuation-in-part of Ser. No.
`289,221, Aug. 12, 1994, abandoned.
`Int. Cl.6
`.............................. C12Q 1/68; C12P 19/34;
`C07H 21102; C07H 21/04
`[52] U.S. Cl ............................. 435/6; 435/91.1; 435/91.2;
`435n.1; 43sn.z; 435n.9; 435/4; 436/548;
`436/500; 530/387.2; 530/388.1; 424/1.11;
`424/88; 536/23.1; 536/24.3; 536124.33
`[58] Field of Search .............................. 435/6, 91.1. 91.2,
`435n.t-7.9, 4; 536/23.1. 24.3-24.33. 548;
`436/500; 424/1.11, 88; 530/387.2, 388.1
`
`[56]
`
`References Cited
`
`U.S. PATENf DOCUMENTS
`
`5,236,844
`
`8/1993 Basset et al ..
`
`111111
`
`11111111111111111111111111111111111111111
`US005753441A
`[11] Patent Number:
`[45] Date of Patent:
`
`5,753,441
`May 19, 1998
`
`FOREIGN PXI'ENT DOCUMENfS
`
`0 518 650
`91109964
`92/00311
`92/11874
`94100764
`95/19369
`
`1211992
`7/1991
`1/1992
`7/1992
`111994
`7/1995
`
`European Pat. Off ..
`WIPO.
`WIPO.
`WIPO.
`WIPO.
`WIPO.
`
`OTHER PUBLICATIONS
`
`Friedman. L.S. et al. (1994). "Confirmation of BRCAl by
`analysis of germline mutations linked to breast and ovarian
`cancer in ten families," Nature Genetics 8:399-404.
`
`(List continued on next page.)
`
`Primary Examiner-W. Gary Jones
`Assistant Examiner-Dianne Rees
`Attorney. Agent, or Firm-Venable, Baetjer. Howard &
`Civiletti. LLP
`
`[57]
`
`ABSTRACT
`
`The present invention relates generally to the field of human
`genetics. Specifically. the present invention relates to meth(cid:173)
`ods and materials used to isolate and detect a human breast
`and ovarian cancer predisposing gene (BRCA1). some
`mutant alleles of which cause susceptibility to cancer. in
`particular breast and ovarian cancer. More specifically. the
`invention relates to germline mutations in the BRCAl gene
`and their use in the diagnosis of predisposition to breast and
`ovarian cancer. The present invention further relates to
`somatic mutations in the BRCA1 gene in human breast and
`ovarian cancer and their use in the diagnosis and prognosis
`of human breast and ovarian cancer. Additionally. the inven(cid:173)
`tion relates to somatic mutations in the BRCA 1 gene in other
`human cancers and their use in the diagnosis and prognosis
`of human cancers. The invention also relates to the therapy
`of human cancers which have a mutation in the BRCAl
`gene. including gene therapy. protein replacement therapy
`and protein mimetics. The invention further relates to the
`screening of drugs for cancer therapy. Finally. the invention
`relates to the screening of the BRCA1 gene for mutations,
`which are useful for diagnosing the predisposition to breast
`and ovarian cancer.
`
`37 Claims, 18 Drawing Sheets
`
`1 tD17S250
`
`t017S800
`
`1
`
`1
`
`1
`
`I
`
`I
`1 BRCA1
`
`1
`
`I
`
`I
`
`' '
`
`01751321
`
`0175855
`
`01751327
`CAI25
`
`RNU2
`
`01751325
`
`GeneDX 1001, pg. 1
`
`

`

`5,753,441
`Page2
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`Chamberlain, J.S., et al. (1993). "BRCA1 Maps Proximal to
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`Feunteun. J .• et al. (1993). "A Breast-Ovarian Cancer Sus(cid:173)
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`Genet. 52:736-742:
`Ford. D .• et al. (1993). 'The risks of cancer in BRCAl
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`Futreal, P.A., et al. (1994). "BRCAl Mutations in Primary
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`dual-specificity phosphatase," Proc. Natl. Acad. Sci. USA
`89:12170-12174.
`Kamb A., et al. (1994). "Localization of the VHR Phos(cid:173)
`phatase Gene and Its Analysis as a Candidate for BRCAl."
`Genomics 23:163-167.
`Kumagai, A. and Dunphy. W.G. (1991). "The cdc25 Protein
`Controls Tyrosine Dephosphorylation of the cdc2 Protein in
`a Cell-Free System," Cell64:903-914.
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`Breast Cancer Families, with Five Chromosome 17q Mark(cid:173)
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`
`GeneDX 1001, pg. 2
`
`

`

`5,753,441
`Page 3
`
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`Goldgar et al. J. of Nat. Cancer lnst. 86: 200-209.1994.
`
`GeneDX 1001, pg. 3
`
`

`

`......
`
`BRCA1
`
`6-8 eM
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`
`

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`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 3 of 18
`
`5,753,441
`
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`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 4 of 18
`
`5,753,441
`
`1
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`c--c------------c-H--c--c----------------c--c
`
`FIG. 5
`
`GeneDX 1001, pg. 7
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 5 of 18
`
`5,753,441
`
`.,..
`co
`
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`
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`GeneDX 1001, pg. 8
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 6 of 18
`
`5,753,441
`
`.... • • • ...
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`
`GeneDX 1001, pg. 9
`
`

`

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`GeneDX 1001, pg. 10
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 8 of 18
`
`5,753,441
`
`mwPabcdefg
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`FIG. 9A
`
`GeneDX 1001, pg. 11
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 9 of 18
`
`5,753,441
`
`2
`
`1
`
`1 2 3 4
`
`f .... ····~~A--
`
`.. -c:---t--~Gt.J..-- --r ........ 1
`
`FIG. 98
`
`GeneDX 1001, pg. 12
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 10 of 18
`
`5,753,441
`
`7
`
`4
`
`PM1
`
`4
`
`5
`
`6
`
`7
`
`8
`
`9
`
`PM7
`
`FIG. 9C
`
`A
`
`G
`
`A
`
`G
`
`A
`
`G
`
`A
`
`G
`
`GeneDX 1001, pg. 13
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 11 of 18
`
`5,753,441
`
`1
`61
`121
`181
`241
`301
`361
`421
`481
`541
`601
`661
`721
`781
`841
`901
`961
`1021
`1081
`1141
`1201
`1261
`1321
`1381
`1441
`1501
`1561
`1621
`1681
`1741
`1801
`1861
`1921
`1981
`2041
`2101
`2161
`2221
`2281
`2341
`2401
`2461
`2521
`2581
`2641
`2701
`2761
`2821
`2881
`2941
`3001
`
`gaggctagagggcaggcactttatggcaaactcaggtagaattcttcctcttccgtctct
`ttccttttacgtcatcggggagactgggtggcaatcgcagcccgagagacgcatggctct
`ttctgccctccatcctctgatgtaccttgatttcgtattctgagaggctgctgcttagcg
`gtagccccttggtttccgtggcaacggaaaagcgcgggaattacagataaattaaaactg
`cgactgcgcggcgtgAGCTCGCTGAGACTTCCTGGACCCCGCACCAGGCTGTGGGGTTTC
`TCAGATAACTGGGCCCCTGCGCTCAGGAGGCCTTCACCCTCTGCTCTGGGTAAAGgtagt
`agagtcccgggaaagggacagggggcccaagtgatgctctggggtactggcgtgggagag
`tggatttccgaagctgacagatgggtattctttgacggggggtaggggcggaacctgaga
`ggcgtaaggcgttgtgaaccctggggaggggggcagtttgtaggtcgcgagggaagcgct
`gaggatcaggaagggggcactgagtgtccgtgggggaatcctcgtgataggaactggaat
`atgccttgagggggacactatgtctttaaaaacgtcggctggtcatgaggtcaggagttc
`cagaccagcctgaccaacgtggtgaaactccgtctctactaaaaatacnaaaattagccg
`ggcgtggtgccgctccagctactcaggaggctgaggcaggagaatcgctagaacccggga
`ggcggaggttgcagtgagccgagatcgcgccattgcactccagcctgggcgacagagcga
`gactgtctcaaaacaaaacaaaacaaaacaaaacaaaaaacaccggctggtatgtatgag
`aggatgggaccttgtggaagaagaggtgccaggaatatgtctgggaaggggaggagacag
`gattttgtgggagggagaacttaagaactggatccatttgcgccattgagaaagcgcaag
`agggaagtagaggagcgtcagtagtaacagatgctgccggcagggatgtgcttgaggagg
`atccagagatgagagcaggtcactgggaaaggttaggggcggggaggccttgattggtgt
`tggtttggtcgttgttgattttggttttatgcaagaaaaagaaaacaaccagaaacattg
`gagaaagctaaggctaccaccacctacccggtcagtcactcctctgtagctttctctttc
`ttggagaaaggaaaagacccaaggggttggcagcgatatgtgaaaaaattcagaatttat
`gttgtctaattacaaaaagcaacttctagaatctttaaaaataaaggacgttgtcattag
`ttcttctggtttgtattattctaaaaccttccaaatcttcaaatttactttattttaaaa
`tgataaaatgaagttgtcattttataaaccttttaaaaagatatatatatatgtttttct
`aatgtgttaaagTTCATTGGAACAGAAAGAAATGGATTTATCTGCTCTTCGCGTTGAAGA
`AGTACAAAATGTCATTAATGCTATGCAGAAAATCTTAGAGTGTCCCATCTGgtaagtcag
`cacaagagtgtattaatttgggattcctatgattatctcctatgcaaatgaacagaattg
`accttacatactagggaagaaaagacatgtctagtaagattaggctattgtaattgctga
`ttttcttaactgaagaactttaaaaatatagaaaatgattccttgttctccatccactct
`gcctctcccactcctctccttttcaacacaatcctgtggtccgggaaagacagggctctg
`tcttgattggttctgcactgggcaggatctgttagatactgcatttgctttctccagctc
`taaavvvvvvvvvvvvvaaatgctgatgatagtatagagtattgaagggatcaatataat
`tctgttttgatatctgaaagctcactgaaggtaaggatcgtattctctgctgtattctca
`gttcctgacacagcagacatttaataaatattgaacgaacttgaggccttatgttgactc
`agtcataacagctcaaagttgaacttattcactaagaatagctttatttttaaataaatt
`attgagcctcatttattttctttttctcccccccctaccctgctagTCTGGAGTTGATCA
`AGGAACCTGTCTCCACAAAGTGTGACCACATATTTTGCAAgtaagtttgaatgtgttatg
`tggctccattattagcttttgtttttgtccttcataacccaggaaacacctaactttata
`gaagctttactttcttcaattaagtgagaacgaaaatccaactccatttcattctttctc
`agagagtatatagttatcaaaagttggttgtaatcatagttcctggtaaagttttgacat
`atattatctttttttttttttgagacaagtctcgctctgtcgcccaggctggagtgcagt
`ggcatgaggcttgctcactgcacctccgcccccgagttcagcgactctvvvvvvvvvvvv
`vtgagatctagaccacatggtcaaagagatagaatgtgagcaataaatgaaccttaaatt
`tttcaacagctactttttttttttttttttgagacagGGKCTTACTCTGTTGTCCCAGCT
`GGAGTACAGWGTGCGATCATGAGGCTTACTGTTGCTTGACTCCTAGGCTCAAGCGATCCT
`ATCACCTCAGTCTCCAAGTAGCTGGACTgtaagtgcacaccaccatatccagctaaattt
`tgtgttttctgtagagacggggtttcgccatgtttcccaggctggtcttgaactttgggc
`ttaacccgtctgcccacctaggcatcccaaagtgctaggattacaggtgtgagtcatcat
`gcctggccagtattttagttagctctgtcttttcaagtcatatacaagttcattttcttt
`taagtttagttaacaaccttatatcatgtattcttttctagcataaagaaagattcgagg
`
`FI-G. 10A
`
`GeneDX 1001, pg. 14
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 12 of 18
`
`5,753,441
`
`3061
`3121
`3181
`3241
`3301
`3361
`3421
`3481
`3541
`3601
`3661
`3721
`3781
`3841
`3901
`3961
`4021
`4091
`4141
`4201
`4261
`4321
`4391
`4441
`4501
`4561
`4621
`4691
`4741
`4801
`4861
`4921
`4981
`5041
`5101
`5161
`5221
`5281
`5341
`5401
`5461
`5521
`5581
`5641
`5701
`5761
`5921
`5881
`5941
`6001
`6061
`
`ccvvvvvvvvvvvvvtgtgatcataacagtaagccatatgcatgtaagttcagttttcat
`agatcattgcttatgtagtttaggtttttgcttatgcagcatccaaaaacaattaggaaa
`ctattgcttgtaattcacctgccattactttttaaatggctcttaagggcagttgtgaga
`ttatcttttcatggctatttgccttttgagtattctttctacaaaaggaagtaaattaaa
`ttgttctttctttctttataatttatagATTTTGCATGCTGAAACTTCTCAACCAGAAGA
`AAGGGCCTTCACAGTGTCCTTTATGTAAGAATGATATAACCAAAAGgtatataatttggt
`aatgatgctaggttggaagcaaccacagtaggaaaaagtagaaattatttaataacatag
`cgttcctataaaaccattcatcagaaaaatttataaaagagtttttagcacacagtaaat
`tatttccaaagttattttcctgaaagttttatgggcatctgccttatacaggtattgvvv
`vvvvvvvvvvggtaggcttaaatgaatgacaaaaagttactaaatcactgccatcacacg
`gtttatacagatgtcaatgatgtattgattatagaggttttctactgttgctgcatctta
`tttttatttgtttacatgtcttttcttattttagtgtccttaaaaggttgataatcactt
`gctgagtgtgtttctcaaacaatttaatttcagGAGCCTACAAGAAAGTACGAGATTTAG
`TCAACTTGTTGAAGAGCTATTGAAAATCATTTGTGCTTTTCAGCTTGACACAGGTTTGGA
`GTgtaagtgttgaatatcccaagaatgacactcaagtgctgtccatgaaaactcaggaag
`tttgcacaattactttctatgacgtggtgataagaccttttagtctaggttaattttagt
`tctgtatctgtaatctattttaaaaaattactcccactggtctcacaccttatttvvvvv
`vvvvvvvvaaaaaatcacaggtaaccttaatgcattgtcttaacacaacaaagagcatac
`atagggtttctcttggtttctttgattataattcatacatttttctctaactgcaaacat
`aatgttttcccttgtattttacagATGCAAACAGCTATAATTTTGCAAAAAAGGAAAATA
`ACTCTCCTGAACATCTAAAAGATGAAGTTTCTATCATCCAAAGTATGGGCTACAGAAACC
`GTGCCAAAAGACTTCTACAGAGTGAACCCGAAAATCCTTCCTTGgtaaaaccatttgttt
`tcttcttctt~tcttcttcttttcttttttttttctttttttttttgagatggagtctt
`gctctgtggcccaggctagaagcagtcctcctgccttagccnccttagtagctgggatta
`caggcacgcgcaccatgccaggctaatttttgtatttttagtagagacggggtttcatca
`tgttggccaggctggtctcgaactcctaacctcaggtgatcvvvvvvvvvvvvvatgatg
`gagatcttaaaaagtaatcattctggggctgggcgtagtagcttgcacctgtaatcccag
`cacttcgggaggctgaggcaggcagataatttgaggtcaggagtttgagaccagcctggc
`caacatggtgaaacccatctctactaaaaatacaaaaattagctgggtgtggtggcacgt
`acctgtaatcccagctactcgggaggcggaggcacaagaattgcttgaacctaggacgcg
`gaggttgcagcgagccaagatcgcgccactgcactccagcctgggccgtagagtgagact
`ctgtctcaaaaaagaaaaaaaagtaattgttctagctgggcgcagtggctcttgcctgta
`atcccagcactttgggaggccaaggcgggtggatctcgagtcctagagttcaagaccagc
`ctaggcaatgtggtgaaaccccatcgctacaaaaaatacaaaaattagccaggcatggtg
`gcgtgcgcatgtagtcccagctccttgggaggctgaggtgggaggatcacttgaacccag
`gagacagaggttgcagtgaaccgagatcacgccaccacgctccagcctgggcaacagaac
`aagactctgtctaaaaaaatacaaataaaataaaagtagttctcacagtaccagcattca
`tttttcaaaagatatagagctaaaaaggaaggaaaaaaaaagtaatgttgggcttttaaa
`tactcgttcctatactaaatgttcttaggagtgctggggttttattgtcatcatttatcc
`tttttaaaaatgttattggccaggcacggtggctcatggctgtaatcccagcactttggg
`aggccgaggcaggcagatcacctgaggtcaggagtgtgagaccagcctggccaacatggc
`gaaacctgtctctactaaaaatacaaaaattaactaggcgtggtggtgtacgcctgtagt
`cccagctactcgggaggctgaggcaggagaatcaactgaaccagggaggtggaggttgca
`gtgtgccgagatcacgccactgcactctagcctggcaacagagcaagattctgtctcaaa
`aaaaaaaaacatatatacacatatatcccaaagtgctgggattacatatatatatatata
`tatatattatatatatatatatatatatatgtgatatatatgtgatatatatataacata
`tatatatgtaatatatatgtgatatatatataatatatatatgtaatatatatgtgatat
`atatatatacacacacacacacatatatatgtatgtgtgtgtacacacacacacacaaat
`tagccaggcatagttgcacacgcttggtagacccagctactcaggaggctgagggaggag
`aatctcttgaacttaggaggcggaggttgcagtgagctgagattgcgccactgcactcca
`gcctgggtgacagagcaggactctgtacaccccccaaaacaaaaaaaaaagttatcagat
`
`FIG. 108
`
`GeneDX 1001, pg. 15
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 13 of 18
`
`5,753,441
`gtgattggaatgtatatcaagtatcagcttcaaaatatgctatattaatacttcaaaaat
`tacacaaataatacataatcaggtttgaaaaatttaagacaacmsaaraaaaaawycmaa
`tcacamatatcccacacattttattattmctmctmcwattattttgwagagmctgggtct
`cacycykttgctwatgctggtctttgaacyccykgccycaarcartcctsctccabcctc
`ccaargtgctggggatwataggcatgarctaaccgcacccagccccagacattttagtgt
`gtaaattcctgggcattttttcaaggcatcatacatgttagctgactgatgatggtcaat
`ttattttgtccatggtgtcaagtttctcttcaggaggaaaagcacagaactggccaa~aa
`ttgcttgactgttctttaccatactgtttagCAGGAAACCAGTCTCAGTGTCCAACTCTC
`TAACCTTGGAACTGTGAGAACTCTGAGGACAAAGCAGCGGATACAACCTCAAAAGACGTC
`TGTCTACATTGAATTGGgtaagggtctcaggttttttaagtatttaataataattgctgg
`attccttatcttatagttttgccaaaaatcttggtcataatttgtatttgtggtaggcag
`ctttgggaagtgaattttatgagccctatggtgagttataaaAaatgtaaaagacgcagt
`tcccaccttgaagaatcttactttaaaaagggagcaaaagaggccaggcatggtggctca
`cacctgtaatcccagcactttgggaggccaaagtgggtggatcacctgaggtcgggagtt
`cgagaccagcctagccaacatggagaaactctgtctgtaccaaaaaataaaaaattagcc
`aggtgtggtggcacataactgtaatcccagctactcgggaggctgaggcaggagaatcac
`ttgaacccgggaggtggaggttgcggtgaaccgagatcgcaccattgcactccagcctgg
`gcaaaaatagcgaaactccatctaaaaaaaaaaaagagagcaaaagaaagamtmtctggt
`tttaamtmtgtgtaaatatgtttttggaaagatggagagtagcaataagaaaaaacatga
`tggattgctacagtatttagttccaagataaattgtactagatgaggaagccttttaaga
`agagctgaattgccaggcgcagtggctcacgcctgtaatcccagcactttgggaggccga
`ggtgggcggatcacctgaggtcgggagttcaagaccagcctgaccaacatggagaaaccc
`catctctactaaaaaaaaaaaaaaaaaaattagccggggtggtggcttatgcctgtaatc
`ccagctactcaggaggctgaggcaggagaatcgcttgaacccaggaagcagaggttgcag
`tgagccaagatcgcaccattgcactccagcctaggcaacaagagtgaaactccatctcaa
`aaaaaaaaaaaaagagctgaatcttggctgggcaggatggctcgtgcctgtaatcctaac
`gctttggaagaccgaggcagaaggattggttgagtccacgagtttaagaccagcctggcc
`aacataggggaaccctgtctctatttttaaaataataatacatttttggccggtgcggtg
`gctcatgcctgtaatcccaatactttgggaggctgaggcaggtagatcacctgaggtcag
`agttcgagaccagcctggataacctggtgaaacccctctttactaaaaatacaaaaaaaa
`aaaaaaattagctgggtgtggtagcacatgcttgtaatcccagctacttgggaggctgag
`gcaggagaatcgcttgaaccagggaggcggaggttacaatgagccaacactacaccactg
`cactccagcctgggcaatagagtgagactgcatctcaaaaaaataataatttttaaaaat
`aataaatttttttaagcttataaaaagaaaagttgaggccagcatagtagctcacatctg
`taatctcagcagtggcagaggattgcttgaagccaggagtttgagaccagcctgggcaac
`atagcaagacctcatctctacaaaaaaatttcttttttaaattagctgggtgtggtggtg
`tgcatctgtagtcccagctactcaggaggcagaggtgagtggatacattgaacccaggag
`tttgaggctgtagtgagctatgatcatgccactgcactccaacctgggtgacagagcaag
`acctccaaaaaaaaaaaaaaaagagctgctgagctcagaattcaaactgggctctcaaat
`tggattttcttttagaatatatttataattaaaaaggatagccatcttttgagctcccag
`gcaccaccatctatttatcataacacttactgttttccccccttatgatcataaattcct
`agacaacaggcattgtaaaaatagttatagtagttgatatttaggagcacttaactatat
`tccaggcactattgtgcttttcttgtataactcattagatgcttgtcagacctctgagat
`tgttcctattatacttattttacagatgagaaaattaaggcacagagaagttatgaaatt
`tttccaaggtattaaacctagtaagtggctgagccatgattcaaacctaggaagttagat
`gtcagagcctgtgctttttttttgtttttgtttttgttttcagtagaaacgggggtctca
`ctttgttggccaggctggtcttgaactcctaacctcaaataatccacccatctcggcctc
`ctcaagtgctgggattacaggtgagagccactgtgcctggcgaagcccatgcctttaacc
`acttctctgtattacatactagcttaactagcattgtacctgccacagtagatgctcagt
`aaatatttctagttgaatatctgtttttcaacaagtacatttttttaacccttttaatta
`agaaaacttttattgatttattttttggggggaaattttttagGATCTGATTCTTCTGAA
`
`6121
`6181
`6241
`6300
`6361
`6421
`6481
`6541
`6601
`6661
`6721
`6781
`6841
`6901
`6961
`7021
`7081
`7141
`7201
`7261
`7321
`7381
`7441
`7501
`7561
`7621
`7681
`7741
`7801
`7861
`7921
`7981
`8041
`8101
`8161
`8221
`8281
`8341
`8401
`8461
`8521
`8581
`8641
`8701
`8761
`8821
`8881
`8941
`9001
`9061
`9121
`
`FIG. 10C
`
`GeneDX 1001, pg. 16
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 14 of 18
`
`5,753,441
`
`9181
`9241
`9301
`9361
`9421
`9481
`9541
`9601
`9661
`9721
`9781
`9841
`9901
`9961
`10021
`10081
`10141
`10201
`10261
`10321
`10381
`10441
`10501
`10561
`10621
`10681
`10741
`10801
`10861
`10921
`10981
`11041
`11101
`11161
`11221
`11281
`11341
`11401
`11461
`11521
`11581
`ll641
`11701
`11761
`11821
`11881
`11941
`12001
`12061
`12121
`12181
`
`GATACCGTTAATAAGGCAACTTATTGCAGgtgagtcaaagagaacctttgtctatgaagc
`tggtattttcctatttagttaatattaaggattgatgtttctctctttttaaaaatattt
`taacttttattttaggttcagggatgtatgtgcagtttgttatataggtaaacacacgac
`ttgggatttggtgtatagatttttttcatcatccgggtactaagcataccccacagtttt
`ttgtttgctttctttctgaatttctccctcttcccaccttcctccctcaagtaggctggt
`gtttctccagactagaatcatggtattggaagaaaccttagagatcatctagtttagttc
`tctcattttatagtggaggaaataccctttttgtttgttggatttagttattagcactgt
`ccaaaggaatttaggataacagtagaactctgcacatgcttgcttctagcagattgttct
`ctaagttcctcatatacagtaatattgacacagcagtaattgtgactgatgaaaatgttc
`aaggacttcattttcaactctttctttcctctgttccttatttccacatatctctcaagc
`tttgtctgtatgttatataataaactacaagcaaccccaactatgttacctaccttcctt
`aggaattattgcttgacccaggtttttttttttttttttttggagacggggtcttgccct
`gttgccaggatggagtgtagtggcgccatctcggctcactgcaatctccaactccctggt
`tcaagcgattctcctgtctcaatctcacgagtagctgggactacaggtatacaccaccac
`gcccggttaattgaccattccatttctttctttctctcttttttttttttttttttgaga
`cagagtcttgctctgttgcccaggctggagtacagaggtgtgatctcacctctccgcaac
`gtctgcctcccaggttgaagccatactcctgcctcagcctctctagtagctgggactaca
`ggcgcgcgccaccacacccggctaatttttgtatttttagtagagatggggtttcaccat
`gttggccaggctggtcttgaactcatgacctcaagtggtccacccgcctcagcctcccaa
`agtgctggaattacaggcttgagccaccgtgcccagcaaccatttcatttcaactagaag
`tttctaaaggagagagcagctttcactaactaaataagattggtcagctttctgtaatcg
`aaagagctaaaatgtttgatcttggtcatttgacagttctgcatacatgtaactagtgtt
`tcttattaggactctgtcttttccctatagTGTGGGAGATCAAGAATTGTTACAAATCAC
`CCCTCAAGGAACCAGGGATGAAATCAGTTTGGATTCTGCAAAAAAGGgtaatggcaaagt
`ttgccaacttaacaggcactgaaaagagagtgggtagatacagtactgtaattagattat
`tctgaagaccatttgggacctttacaacccacaaaatctcttggcagagttagagtatca
`ttctctgtcaaatgtcgtggtatggtctgatagatttaaatggtactagactaatgtacc
`tataataagaccttcttgtaactgattgttgccctttcgcttttttttttgtttgtttgt
`ttgtttttttttgagatggggtctcactctgttgcccaggctggagtgcagtgatgcaat
`cttggctcactgcaacctccacctccaaaggctcaagctatcctcccacttcagcctcct
`gagtagctgggactacaggcgcatgccaccacacccggttaattttttgtggttttatag
`agatggggtttcaccatgttaccgaggctggtctcaaactcctggactcaagcagtctgc
`ccacttcagcctcccaaagtgctgcagttacaggcttgagccactgtgcctggcctgccc
`ttt&cttttaattggtgtatttgtgtttcatcttttacctactggtttttaaatataggg
`agtggtaagtctgtagatagaacagagtattaagtagacttaatggccagtaatctttag
`agtacatcagaaccagttttctgatggccaatctgcttttaattcactcttagacgttag
`agaaataggtgtggtttctgcatagggaaaattctgaaattaavvvvvvvvvvvvvgatc
`ctaagtggaaataatctaggtaaataggaattaaatgaaagagtatgagctacatcttca
`gtatacttggtagtttatgaggttagtttctctaatatagccagttggttgatttccacc
`tccaaggtgtatgaagtatgtatttttttaatgacaattcagtttttgagtaccttgtta
`tttttgtatattttcagCTGCTTGTGAATTTTCTGAGACGGATGTAACAAATACTGAACA
`TCATCAACCCAGTAATAATGATTTGAACACCACTGAGAAGCGTGCAGCTGAGAGGCATCC
`AGAAAAGTATCAGGGTAGTTCTGTTTCAAACTTGCATGTGGAGCCATGTGGCACAAATAC
`TCATGCCAGCTCATTACAGCATGAGAACAGCAGTTTATTACTCACTAAAGACAGAATGAA
`TGTAGAAAAGGCTGAATTCTGTAATAAAAGCAAACAGCCTGGCTTAGCAAGGAGCCAACA
`TAACAGATGGGCTGGAAGTAAGGAAACATGTAATGATAGGCGGACTCCCAGCACAGAAAA
`AAAGGTAGATCTGAATGCTGATCCCCTGTGTGAGAGAAAAGAATGGAATAAGCAGAAACT
`GCCATGCTCAGAGAATCCTAGAGATACTGAAGATGTTCCTTGGATAACACTAAATAGCAG
`CATTCAGAAAGTTAATGAGTGGTTTTCCAGAAGTGATGAACTGTTAGGTTCTGATGACTC
`ACATGATGGGGAGTCTGAATCAAATGCCAAAGTAGCTGATGTATTGGACGTTCTAAATGA
`GGTAGATGAATATTCTGGTTCTTCAGAGAAAATAGACTTACTGGCCAGTGATCCTCATGA
`
`FIG. 100
`
`GeneDX 1001, pg. 17
`
`

`

`U.S. Patent
`
`May 19, 1998
`
`Sheet 15 of 18
`
`5,753,441
`
`12241
`12301
`12361
`12421
`12481
`12541
`12601
`12661
`12721
`12781
`12841
`12901
`12961
`13021
`13081
`13141
`13201
`13261
`13321
`13381
`13441
`13501
`13561
`13621
`13681
`13741
`13801
`13861
`13921
`13981
`14041
`14101
`14161
`14221
`14281
`14341
`14401
`l4461
`14521
`14581
`14641
`14701
`14761
`14821
`14881
`14941
`15001
`15061
`15121
`15181
`15241
`
`GGCTTTAATATGTAAAAGTGAAAGAGTTCACTCCAAATCAGTAGAGAGTAATATTGAAGG
`CCAAATATTTGGGAAAACCTATCGGAAGAAGGCAAGCCTCCCCAACTTAAGCCATGTAAC
`TGAAAATCTAATTATAGGAGCATTTGTTACTGAGCCACAGATAATACAAGAGCGTCCCCT
`CACAAATAAATTAAAGCGTAAAAGGAGACCTACATCAGGCCTTCATCCTGAGGATTTTAT
`CAAGAAAGCAGATTTGGCAGTTCAAAAGACTCCTGAAATGATAAATCAGGGAACTAACCA
`AACGGAGCAGAATGGTCAAGTGATGAATATTACTAATAGTGGTCATGAGAATAAAACAAA
`AGGTGATTCTATTCAGAATGAGAAAAATCCTAACCCAATAGAATCACTCGAAAAAGAATC
`TGCTTTCAAAACGAAAGCTGAACCTATAAGCAGCAGTATAAGCAATATGGAACTCGAATT
`AAATATCCACAATTCAAAAGCACCTAAAAAGAATAGGCTGAGGAGGAAGTCTTCTACCAG
`GCATATTCATGCGCTTGAACTAGTAGTCAGTAGAAATCTAAGCCCACCTAATTGTACTGA
`ATTGCAAATTGATAGTTG'l'TCTAGCAGTGAAGAGATAAAGAAAAAAAAGTACAACCAAAT
`GCCAGTCAGGCACAGCAGAAACCTACAACTCATGGAAGGTAAAGAACCTGCAACTGGAGC
`CAAGAAGAGTAACAAGCCAAATGAACAGACAAGTAAAAGACATOACAGCGATACTTTCCC
`AGAGCTGAAGTTAACAAATGCACCTGG%TCTTTTACTAAGTGTTCAAATACCAGTGAACT
`TAAAGAATTTGTCAATCCTAGCCTTCCAAGAGAAGAAAAAGAAGAGAAACTAGAAACAGT
`TAAAGTGTCTAATAATGCTGAAGACCCCAAAGATCTCATGTTAAGTGGAGAAAGGGTTTT
`GCAAACTGAAAGATCTGTAGAGAGTAGCAGTATTTCA%TGGTACCTGGTACTGATTATGG
`CAt!TCAGGAAAGTATCTCGTTACTGGAAGTTAGCACTCTAGGGAAGGCAAAAACAGAACC
`AAATAAATGTGTGAGTCAGTGTGCAGCATTTGAAAACCCCAAGGGACTAATTCATGGTTG
`TTCCAAAGATAATAGAAATGACACAGAAGGCTTTAAGTATCCATTGGGACATGAAGTTAA
`CCACAGTCGGGAAACAAGCATAGAAATGGAAGAAAGTGAACTTGATGCTCAGTATTTGCA
`GAATACATTCAAGGTTTCAAAGCGCCAGTCATTTGCTCCGTTTTCAAATCCAGGAAATGC
`AGAAGAGGAATGTGCAACATTCTCTGCCCACTCTGGGTCCTTAAAGAAACAAAGTCCAAA
`AGTCACTTTTGAATGTGAACAAAAGGAAGAAAATCAAGGAAAGAATGAGTCTAATATCAA
`GCCTGTACAGACAGTTAATATCACTGCAGGCTTTCCTGTGGTTGGTCAGAAAGATAAGCC
`AGTTGATAATGCCAAATGTAGTATCAAAGGAGGCTCTAGGTTTTGTCTATCATCTCAGTT
`CAGAGGCAACGAAACTGGACTCATTACTCCAAATAAACATGGACTTTTACAAAACCCATA
`TCGTATACCACCACTTTTTCCCATCAAGTCATTTGTTAAAACTAAATGTAAGAAAAATCT
`GCTAGAGGAAAACTTTGAGGAACATTCAATaTCACCTGAAAGAGAAATGGGAAATGAGAA
`CATTCCAAGTACAGTGAGCACAATTAGCCGTAATAACATTAGAGAAAATGTTTTTAAAGA
`AGCCAQCTCAAGCAATATTAATGAAGTAGGTTCCAGTACTAATGAAGTGGGCTCCAGTAT
`TAATGAAATAGGTTCCAGTGATGAAAACATTCAAGCAGAACTAGGTAGAAACAGAGGGCC
`AAAATTGAATGCTATGCTTAGATTAGGGGTTTTGCAACCTGAGGTCTATAAACAAAGTCT
`TCCTGGAAGTAATTGTAAGCATCCTGAAATAAAAAAGCAAGAATATGAAGAAGTAGTTCA
`GACTGTTAATACAGATTTCTCTCCATATCTGATTTCAGATAACTTAGAACAGCCTATGGG
`AAGTAGTCATGCATCTCAGGTTTGTTCTGAGACACCTGATGACCTGTTAGATGATGGTGA
`AATAAAGGAAG