■ 基本信息
別名: | pCDFDuet1 |
啟動子: | T7 |
復(fù)制子: | CDF |
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: | 3781bp |
質(zhì)粒標(biāo)簽: | N-6×His,C-S |
原核抗性: | Str |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，有氧，LB |
表達(dá)宿主: | BL21(DE3) |
誘導(dǎo)方式: | IPTG或乳糖及其類似物 |
5'測序引物: | ACYCDuetUP1：GGATCTCGACGCTCTCCCT DuetUP2:TTGTACACGGCCGCATAATC |
3'測序引物:
| DuetDOWN1:GATTATGCGGCCGTGTACAA T7-ter:TGCTAGTTATTGCTCAGCGG
|
備注: | 同時表達(dá)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(dá) |
片段類型: | ORF
|
片段物種: | 多框空載
|
原核抗性: | Str |
真核抗性: |
|
熒光標(biāo)記: |
|
■ 質(zhì)粒簡介
pCDFDuet-1質(zhì)粒是一種原核表達(dá)載體， N端含有一個6×His標(biāo)簽，C端含有一個S標(biāo)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(dá)由宿主細(xì)胞提供的T7 RNA聚合酶誘導(dǎo)，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CloDF13復(fù)制因子、lacI基因和鏈霉素抗性基因?？梢允惯@個pETDuet-1載體在一個適當(dāng)?shù)乃拗骶甑腸oexpression 4目標(biāo)基因插入MCS1的基因可以使用ACYCDuetUP1引物和DuetDOWN1引物進(jìn)行測序。插入MCS2的基因可以用DuetUP2引物和T7進(jìn)行排序。
pCDFDuet-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 CloDF13-derived CDF replicon, lacI gene and streptomycin/spectinomycin resistance gene (SmR). This vector can be used in combination with pACYCDuet?-1 , pRSFDuet?-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)粒只保證關(guān)鍵序列正確，不保證表達(dá)效果。
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請下載：
ZK324pCDFDuet-1大腸雙表達(dá)質(zhì)粒.txt
