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US4401759A Expired / Lapsed US ⏱️ 0.0 years remaining

Detection and isolation of glucagon mRNA using a synthetic oligodeoxynucleotide

Assignee: Individual
•
Filing: 1982-04-08
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Granted: 1983-08-30
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Expires: 2002-04-08
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Abstract

Abstract A synthetic oligodeoxynucleotide complementary to glucagon mRNA and a method of using it to detect and isolate glucagon mRNA and cDNA from human and rabbit pancreas. A unique 14 base oligodeoxynucleotide dTTCATCAGCCACTG wherein T represents thymine, G represents guanine, A represents adenine, and C represents cytosine and where at least 13 of the 14 nucleotides are as indicated (one of those indicated may be replaced by one of the other three mentioned nucleotides) has been found to be complementary to glucagon mRNA. To isolate glucagon mRNA, total RNA is first extracted from human or rabbit pancreas and A+ RNA (mRNA-poly A) is isolated from the total RNA. The A+ RNA is then treated with the oligodeoxynucleotide, and the resulting hybridized RNA is enzymatically converted to glucagon mRNA which can then be purified, copied into glucagon cDNA and used in a conventional manner to produce glucagon by cloning technique.

Inventors

  • 👤 Harvey Rubin
  • 👤 Richard H. Tullis

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-01 19:12
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/605; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333

Claims Analysis

Total Claims: 7
Claim 1
Independent Claim 68 words
The method of isolating glucagon mRNA from human and rabbit pancreas which comprises the steps of: extracting total RNA from rabbit or human pancreas; isolating mRNA-poly A from said total RNA; and hybridizing said mRNA-poly A with a 14 base oligodeoxynucleotide dTTCATCAGCCACTG, wherein A represents adenine, T represents thymine, G represents guanine and C represents cytosine, and wherein at least 13 of the 14 nucleotides are as indicated.
Claim 2
Independent Claim 68 words
The method of isolating glucagon mRNA from human and rabbit pancreas which comprises the steps of: extracting total RNA from rabbit or human pancreas; isolating mRNA-poly A from said total RNA; and hybridizing said mRNA-poly A with a 14 base oligodeoxynucleotide dTTCATCAGCCACTG, wherein A represents adenine, T represents thymine, G represents guanine and C represents cytosine, and wherein at least 13 of the 14 nucleotides are as indicated.
Claim 3
Dependent Claim 18 words
The method according to claim 1 wherein said hybridization temperature range is from about 0° to 50° C.
Claim 4
Dependent Claim 32 words
The method according to claim 1 further including the steps of copying said hybridized mRNA into complementary DNA and isolating said complementary DNA whereby substantially only cDNA resulting from glucagon mRNA remains.
Claim 5
Independent Claim 82 words
The method of isolating glucagon mRNA from human pancreas which comprises the steps of: separating pancreas islets from fresh human pancreas; extracting total RNA from said islets by guanidine hydrochloric acid extraction; isolating mRNA-poly A from said total RNA on oligo-dT-cellulose; hybridizing said mRNA-poly A in a sodium phosphate buffer with the 14 base oligodeoxynucleotide dTTCATCAGCCAACTG, wherein A represents adenine, T represents thymine, G represents guanine and C represents cytosine, and wherein at least 13 of the 14 nucleotides are as indicated.
Claim 6
Independent Claim 82 words
The method of isolating glucagon mRNA from human pancreas which comprises the steps of: separating pancreas islets from fresh human pancreas; extracting total RNA from said islets by guanidine hydrochloric acid extraction; isolating mRNA-poly A from said total RNA on oligo-dT-cellulose; hybridizing said mRNA-poly A in a sodium phosphate buffer with the 14 base oligodeoxynucleotide dTTCATCAGCCAACTG, wherein A represents adenine, T represents thymine, G represents guanine and C represents cytosine, and wherein at least 13 of the 14 nucleotides are as indicated.
Claim 7
Dependent Claim 22 words
The method according to claim 4 further including the step of analyzing the isolated mRNA-poly A prior to hybridizing said mRNA-poly A.

References Cited (Backward Citations)

US3689475A Cited
US4237224A Cited
US4321365A Cited
US4224408A Cited
US4293652A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1986-08-29 1987-02-28 1987-08-30 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1990-08-29 1991-02-28 1991-08-30 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1994-08-29 1995-02-28 1995-08-30 $7700.00 Window Closed / Lapsed

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