| Project/Area Number |
04557057
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
TANAKA Koichi Kyoto University, school of medicine assistant professor, 医学部, 助教授 (20115877)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIZAKI Kanji Radiation Biology Research Center assistant professor, 放射線生物研究センター, 助教授 (70111987)
HONDA Kazuo School of Medicine assistant, 医学部, 助手 (00209321)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥12,900,000 (Direct Cost: ¥12,900,000)
Fiscal Year 1993: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1992: ¥8,700,000 (Direct Cost: ¥8,700,000)
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| Keywords | partial liver transplantation / liver function / aspiration cytology / RT-PCR / PT-PCR / 肝再生 |
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| Research Abstract |
To evaluate the liver function and the regeneration ability after clinical living related donor partial liver transplantation, the method for the determination of the amount of the mRNA of albumin in the hepatocytes contained in the small samples obtained by percutaneous fine needle aspiration biopsy was developed using competitive RT-PCR.The primer for site A was selected from the sequence between exon 4 and exon 5 and the primer for site B was selected from the sequence between exon 11 and exon 12 of the rat albumin gene. The primers introduced with a point mutation, which generate a new restriction site, were synthesized for the site A and the site B.The PCR product from the cDNA obtained by reverse transcriptase using these primers with the point mutation was used as the competitor (internal control), which was recognized by electrophoresis after digestion with the specific endonuclease. Total nucleic acids were extracted from the small amount of the samples of rat liver by Iso Quick^<TM>. The cDNAs were translated from mRNAs using Super Script Preamplification Kit^<TM> and amplified with the genomic DNA and the various amounts of the competitor by PCR.The quantitation of the initial amounts of the mRNA for albumin in the sample was achieved by the determination of the ratio of these products by dencitometry. This method may provide a rapid way to quantitate the amount of the mRNA of albumin in the very small amount of the clinical samples obtained by fine needle aspiration biopsy.
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