Oligonucleotide probes for detection of .alpha.-amylase genes

• US 4,917,999 A
• Filed: 09/02/1986
• Issued: 04/17/1990
• Est. Priority Date: 09/02/1986
• Status: Expired due to Fees

• Current Assignee: Miles, Inc. (Bayer AG)
• Original Assignee: Miles, Inc. (Bayer AG)
• Inventors: Byng, Graham, Wollweber, Karen L.
• Primary Examiner(s): Griffin, Ronald W.
• Assistant Examiner(s): CRANE, LAWRENCE E
• Application Number:
  US06/902,905
• Time in Patent Office: 1,323 Days
• Field of Search: 935/77, 935/78, 435/5, 435/6, 435/7, 536/22, 536/28, 536/29
• US Class Current: 435/6.13
• CPC Class Codes: C12N 9/2417 from microbiological source

Abstract

Methods and compositions are provided for detecting a gene encoding αamylase from a nucleic acid sample suspected of containing said gene. Said methods utilize oligonucleotide probes of consensus sequences deduced from highly conserved DNA sequences of various Bacillus species.

Claims

1. An oligonucleotide probe for use in the detection of a gene encoding α

-amylase in an organism suspected of carrying the gene, comprising;

an oligonucleotide having a base sequence chosen from the sequences,

(a) GAI GGI TTI CGI ITT GAT GCC GIC AAI CAT ATT

wherein I is inosine;

or

(b) GTT ACA TTT GTT GAA AAT CAT GAC ACA

which oligonucleotides are capable of hybridization, under hybridization conditions, with a second base sequence present in the organisms Bacillus stearothermophilus, Bacillus subtilis NA64, Bacillus amuloliquefaciens and Bacillus licheniformis, which second base sequence is capable of encoding the aminoacid sequence asp-gly-phe-arg-leu-asp-ala-val-lys-his-ile.

3. A method for detecting the presence of a gene encoding the enzyme α

-amylase from an organism suspected of containing said gene comprising contacting the oligonucleotide probe of claim 1 with a DNA sample from said organism under conditions favorable to hybridization between said probe and said sample and determining the presence of hybridized probe.

2. The oligonucleotide probe comprising the sequence:

(a) GAI GGI TTI CGI ITT GAT GCC GIC AAI CAT ATT

wherein I is inosine;

or the sequence

(b) GTT ACA TTT GTT GAA AAT CAT GAC ACA.

5. A method for detecting the presence of a gene encoding the enzyme α

amylase from an organism suspected of containing said gene comprising contacting the oligonucleotide probe of claim 2 with a DNA sample from said organism under conditions favorable to hybridization between said probe and said sample and determining the presence of hybridized probe.

4. A method for detecting the presence of a gene encoding the enzyme α

-amylase from an organism suspected of containing said gene comprising contacting an oligonucleotide probe of the sequence;

GAI GGI TTI CGI ITT GAT GCC GIC AAI CAT ATT

wherein I is inosine with a DNA sample from said organism under conditions favorable to hybridization between said probe and said sample and determining the presence of hybridized probe.

6. A method for detecting the presence of a gene encoding the enzyme α

-amylase from an organism suspected of containing said gene comprising contacting an oligonucleotide probe of the sequence;

GTT ACA TTT GTT GAA AAT CAT GAC ACA

with a DNA sample from said organism under conditions favorable to hybridization between said probe and said sample and determining the presence of hybridized probe.