Chiba/Flagella/Making FliC-his
From 2007.igem.org
(Difference between revisions)
| Line 5: | Line 5: | ||
#GelCheck | #GelCheck | ||
| + | |||
| + | *design primer | ||
| + | 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) | ||
| + | **pUC19-FliCをpUC19-FliC-vectorに | ||
| + | [PCR表] | ||
[混ぜ表] | [混ぜ表] | ||
*Template1μL | *Template1μL | ||
| Line 10: | Line 15: | ||
*10×buffer10μL | *10×buffer10μL | ||
*dNT | *dNT | ||
| - | + | *design insert | |
| - | *design | + | **six histidine loop peptide ("His-loop") |
| - | * | + | **アミノ酸配列-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' | |
| - | *six histidine loop peptide ("His-loop") | + | |
| - | *アミノ酸配列-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' | + | |
Revision as of 03:24, 27 October 2007
- pUC19 vectorにFliCをコードする遺伝子を挿入→pUC19-FliC(島田さんから)
- エキサイトPCR
- design insert(ヒスチジンタグ)
- pUC19-FliC vector とinsertオリゴをライゲーション.
- GelCheck
- design primer
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)
- pUC19-FliCをpUC19-FliC-vectorに
[PCR表] [混ぜ表]
- Template1μL
- Primer(FとR)5μL
- 10×buffer10μL
- dNT
- design insert
- six histidine loop peptide ("His-loop")
- アミノ酸配列-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'
