| Project/Area Number |
13470214
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Embryonic/Neonatal medicine
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| Research Institution | Tokyo Women's Medical University |
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Principal Investigator |
NAKANISHI Toshio Tokyo Women's Medical University, Dep. Of Medicine, Associate Professor, 医学部, 助教授 (90120013)
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| Co-Investigator(Kenkyū-buntansha) |
AIBA Sumi Tokyo Women's Medical University, Dep. Of Medicine, Assistant Professor, 医学部, 講師 (80138891)
MATSUOKA Rumiko Tokyo Women's Medical University, Dep. Of Medicine, Assistant Professor, 医学部, 講師 (50120051)
HAGIWARA Nobuhisa Tokyo Women's Medical University, Dep. Of Medicine, Associate Professor, 医学部, 助教授 (00180802)
HAGANA Emiko Tokyo Women's Medical University, Dep. Of Medicine, Assistant Professor, 医学部, 助手
NAKAZAWA Makoto Tokyo Women's Medical University, Dep. Of Medicine, Professor, 医学部, 教授 (10075567)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2002: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥11,200,000 (Direct Cost: ¥11,200,000)
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| Keywords | Ductus Arteriosus / K channel / Oxygen / Cloning of K channel / クローニング / 遺伝子 |
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| Research Abstract |
Establishment of cDNA of ductus arteriosus isolated from pig :
Ductal tissue, aorta, pulmonary artery was isolated from 140 newborn pigs immediately after birth. Total RNA and then poly A RNA was isolated from the ductus, aorta, and pulmonary artery. cDNA was made from the RNA using RT-PCR. The cDNA was inserted into vector and cDNA library was established. The length of the inserted DNA was between 1.7 to 2.7 kb.
Cloning Kv using the cDNA library :
The cDNA library of pig ductus arteriosus was screened using Kv probes obtained from rat, mouse, and human beings. Kv 1, Kv 2, and Kv9.3 were cloned from the ductus arteriosus.
Screening of the cDNA library using homology-based PCR :
Primer was made using conserved part of Kv genes and PCR was performed using cDNA as a template. PCR products were compared between the ductus, aorta, and pulmonary artery. PCR products amplified only in the ductus was chosen and cloned. Kv families expressed in the ductus were cloned and they belong to 1.5, 2.1 and 2.2 family.
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