書誌事項
- タイトル別名
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- Direct Expression of the Pig Testicular 3α/β (20β)-Hydroxysteroid Dehydrogenase cDNA Using Baculovirus Expression System
- ブタ精巣3α/β(20β)-ヒドロキシステロイドデヒドロゲナーゼcDNAのバキュロウイルス発現系を利用したタンパク質直接発現
- ブタ セイソウ 3アルファ ベータ 20ベータ ヒドロキシステロイドデヒドロ
- Direct Expression of the Pig Testicular 3α/β (20β)-Hydroxysteroid Dehydrogenase cDNA Using Baculovirus Expression System
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The direct expression of the pig testicular 3α/β (20β)-hydroxysteroid dehydrogenase (HSD) cDNA using a baculovirus expression system was investigated. A minimum essential cDNA containing the coding region of 3α/β (20β)-HSD was amplified by polymerase chain reaction (PCR) to extend the DNA linker including the Bam HI restriction site in the both ends of the cDNA. As the template, 3α/β (20β)-HSD cDNA, pBS 52, which was subcloned to Bluescript II was used. The PCR fragment was ligated to Bam HI-cut transfer plasmid (pBlueBac III). Recombinant transfer plasmid (pBlueBac-20β) constructed was cotransfected into Spodoptera frugiperda Sf-9 cells with the baculovirus. After transfection, the recombinant virus was detected by the plaque assay with color selection. The expression of 3α/β (20β)-HSD cDNA was detected by Western blotting and enzyme assay. The expressed protein showed the same molecular weight and immunochemical cross-reaction to the native enzyme. Furthermore, it had 3-keto reductase activity of 3α/β (20β)-HSD for 5α-dihydrotestosterone and 20-keto reductase activity for 17α-hydroxyprogesterone in the presence of NADPH.
収録刊行物
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- 薬学雑誌
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薬学雑誌 115 (4), 344-350, 1995
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390001206155016064
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- NII論文ID
- 110003648954
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- NII書誌ID
- AN00284903
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- ISSN
- 13475231
- 00316903
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- NDL書誌ID
- 3620412
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- JaLC
- NDL
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