| Project/Area Number |
01570386
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Hamamatsu University, School of Medicine |
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Principal Investigator |
KANEKO Eizo Hamamatsu University, School of Medicine, Professor, 医学部, 教授 (10010414)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Masafumi Hamamatsu University, School of Medicine, Research Associate, 医学部, 助手 (50179256)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1990: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Peptic ulcer / Gastrin mRNA / Northern blot hybridization / Polymerase chain reaction / Polymerase Chain Reaction法 / ガストリン / mRNA / 合成DNAプロ-ブ |
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| Research Abstract |
We developed a plain polymerase chain reaction (PCR) technique sensitive enough to detect gastrin mRNA in gastric antral mucosa obtained from the patients of peptic ulcer by endoscopic biopsy. From only 1 mug of total cellular RNA we can detect gastrin mRNA.
Gastric antral mucosas were obtained by endoscopic biopsy in the patients with peptic ulcers. We can obtain 20 mug of total cellular RNA form 40 mg of gastric antral mucosa. We chose beta-actin as our internal control because, as a major component of the cytoskeletal system, it is transcribed in most eukaryotic cells. Northern blot hybridizations for gastrin mRNA and beta-actin were carried out using synthetic oligodeoxyribonucleotides. Beta-actin mRNAs (2.0 kb) were detected in all specimens examined, but gastrin mRNAs (0.5 kb) was detected in half of them by Northern hybridization. We used 1 mug of total RNA in our gastrin PCR assay. Total cellular RNA was treated with DNase to remove any contaminating DNA. We also chose beta-actin as internal control of amplification. Gastrin and beta-actin transcripts can be simultaneously amplified from gastric antral mucosa. The ratio of PCR-amplified gastrin to beta-actin signal was well correlated with the ratio of gastrin to beta-actin signal in Northern blot hybridization.
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