Chiba/Flagella/Making FliC-his

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  • pUC19 vectorにFliCをコードする遺伝子を挿入→pUC19-FliC
  • FliCの可変領域のうち、フィラメントの外側部分から５か所を選び、プライマーを設計(207,228,240,252,267)し、エキサイトPCR→pUC19-FliC vector
  • EcoRⅠSite,2×Ser,6×His,2×Ser,NcoⅠ siteをコードするオリゴをデザイン。

(six histidine loop peptide ("His-loop")(containing imidazole side chain), -Ser-Ser-His-His-His-His-His-His-Ser-Ser,encoded by the oligo 5'-TCT-TCT-CAT-CAT-CAT-CAT-CAT-CAT-TCC-TCC-3' and anti oligo 5'-GGA-GGA-ATG-ATG-ATG-ATG-ATG-ATG-AGA-AGA-3')

  • pUC19-FliC vector と6×Hisオリゴをライゲーション.
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