APC gene and nucleic acid probes derived therefrom

• US 5,352,775 A
• Filed: 08/08/1991
• Issued: 10/04/1994
• Est. Priority Date: 01/16/1991
• Status: Expired

• Current Assignee: Zeneca Incorporated (Astrazeneca PLC), Japanese Foundation For Cancer Research Cancer Institute, Johns Hopkins University, University of Utah (State of Utah)
• Original Assignee: Cancer Institute, IMPERIAL CHEMICAL INDUSTRIES, Johns Hopkins University, University of Utah (State of Utah)
• Inventors: Markham, Alexander F., Hedge, Philip J., Nakamura, Yusuke, Vogelstein, Bert, Albertsen, Hans, Kinzler, Kenneth, Thliveris, Andrew, Groden, Joanna, Anand, Rakesh, Joslyn, Geoff, Carlson, Mary, White, Raymond L.
• Primary Examiner(s): Hill, Jr., Robert J.
• Assistant Examiner(s): Carlson, K. Cochrane
• Application Number:
  US07/741,940
• Time in Patent Office: 1,153 Days
• Field of Search: 435/6, 435/810, 935/8, 935/78, 536/27
• US Class Current: 536/23.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/172 Haplotypes
• CPC Class Codes: Y10S 530/828 Cancer

Abstract

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

Claims

1. A cDNA molecule having the nucleotide sequence shown in SEQ ID NO:

1 or its complement.

2. An isolated DNA molecule having the nucleotide sequence shown in SEQ ID NO:

1 or its complement.

3. A cDNA molecule which encodes a protein having the amino acid sequence shown in FIG. 3 or 7 (SEQ ID NO:

7 or

     2).

4. An isolated DNA molecule which encodes a protein having the amino acid sequence shown in FIG. 3 or 7 (SEQ ID NO:

7 or

     2).

5. A nucleic acid probe complementary to all or part of human wild-type APC gene coding sequences or the complement of said sequence such that said probe selectively hybridizes under stringent conditions to said APC gene or identifies endogenous, random modifications in said APC gene.

6. The nucleic acid probe of claim 5 which hybridizes to all or part of an exon selected from the group consisting of:

(1) nucleotides 822 to 930;

(2) nucleotides 931 to 1309;

(3) nucleotides 1406 to 1545; and

(4) nucleotides 1956 to 2256.

7. A set of probes useful for detecting alteration of wild-type APC genes comprising a plurality of nucleic acid probes wherein said set is complementary to all nucleotides of the APC gene coding sequences as shown in SEQ ID NO:

1 or the complement of said sequences.

8. A pair of signal stranded DNA primers for determination of a nucleotide sequence of an APC gene by polymerase chain reaction, the sequence of said primers being derived from said APC gene, wherein the use of said primers in a polymerase chain reaction results in synthesis of DNA having all or part of the sequence shown in FIG. 7 (SEQ ID NO:

1).

9. The pair of primers of claim 8 which have restriction enzyme sites at each 5'"'"' end.

10. The pair of primers of claim 9 having sequences complementary to all or part of one or more APC introns.