■ 基本信息
別名: | pRSFDuet1 |
啟動子: | T7 |
復制子: | RSF |
終止子: | T7 terminator |
質(zhì)粒分類: | 大腸系列質(zhì)粒；大腸其它質(zhì)粒；大腸雙框質(zhì)粒 |
質(zhì)粒大小: | 3829bp |
質(zhì)粒標簽: | N-6×His，C-S |
原核抗性: | Kan |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度，37℃，有氧，LB |
表達宿主: | BL21(DE3)等大腸桿菌 |
誘導方式: | IPTG或乳糖及其類似物 |
5'測序引物: | ACYCDuetUP1-F(GGATCTCGACGCTCTCCCT) DuetUP2-F( TTGTACACGGCCGCATAATC) |
3'測序引物:
| DuetDOWN1-R( GATTATGCGGCCGTGTACAA) T7-ter-R( TGCTAGTTATTGCTCAGCGG) |
備注: | 同時表達兩個外源蛋白 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 大腸桿菌 |
質(zhì)粒用途: | 蛋白表達 |
片段類型: | ORF
|
片段物種: | 多框空載
|
原核抗性: | Kan |
真核抗性: |
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熒光標記: |
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■ 質(zhì)粒簡介
pRSFDuet-1質(zhì)粒是一種原核表達載體， N端含有一個6×His標簽，C端含有一個S標簽。該質(zhì)粒含有多個常用的酶切位點，便于不同基因克隆。表達由宿主細胞提供的T7 RNA聚合酶誘導，目的基因被克隆到質(zhì)粒載體上，受噬菌體強轉(zhuǎn)錄及翻譯信號控制。
該質(zhì)粒還攜帶有RSF復制因子、lacI基因和氯霉素抗性基因?？梢允惯@個pETDuet-1載體在一個適當?shù)乃拗骶甑腸oexpression 4目標基因插入MCS1的基因可以使用ACYCDuetUP1引物和DuetDOWN1引物進行測序。插入MCS2的基因可以用DuetUP2引物和T7進行排序。
pRSFDuet-1 is designed for the coexpression of two target ORFs. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7lac promoter and ribosome binding site (rbs). The vector also carries the RSF1030-derived RSF replicon, lacI gene and kanamycin resistance gene (KnR). This vector can be used in combination with pACYCDuet?-1 , pCDFDuet?-1 , and pETDuet?-1 in an appropriate host strain for the coexpression of 4 to 8 target proteins. ORFs inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer . ORFs inserted into MCS2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer . Unique sites are shown on the circle map.
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請下載：
ZK1686 pRSFDuet-1大腸雙表達質(zhì)粒.txt
