■ 基本信息
別名: | pTWINI |
啟動(dòng)子: | T7 |
復(fù)制子: | M13 |
質(zhì)粒分類: | 大腸桿菌載體；蛋白表達(dá)質(zhì)粒 |
質(zhì)粒大小: | 7375bp |
原核抗性: | Amp |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，有氧，LB |
表達(dá)宿主: | 大腸桿菌 |
誘導(dǎo)方式: | IPTG或乳糖及其類似物 |
備注: | 表達(dá)質(zhì)粒 (大腸桿菌融合標(biāo)簽自剪切表達(dá)載體) |
■ 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: | 空載體 |
原核抗性: | Amp |
真核抗性: |
|
熒光標(biāo)記: |
|
■ 質(zhì)粒簡(jiǎn)介
pTWIN1 is an E. coli plasmid cloning vector designed for recombinant protein expression, labeling, and cyclization using the IMPACT-TWIN Kit (1). It contains the pMB1 origin of replication from pBR322 and is maintained at a similar copy number to pBR322; in addition, pTWIN1 also contains an M13 origin of replication.
The multiple cloning site (MCS) is positioned to allow translational fusion of an intein tag to the N-terminus, C-terminus, or both, of the cloned target protein. The mini-inteins encoded by pTWIN1, the Ssp DnaB intein and the Mxe GyrA intein, cleave the peptide bond at their C- and N-termini, respectively (1-4). The chitin binding domain (CBD) from B. circulans, fused to each intein, facilitates purification of the intein-target protein precursor.
Transcription of the intein tags is controlled by the inducible T7 promoter, requiring E. coli strains containing integrated copies of the T7 RNA polymerase gene [e.g. BL21(DE3)] for expression. Basal expression from the T7 promoter is minimized by the binding of the Lac repressor, encoded by the lacI gene, to the lac operator immediately downstream of the T7 promoter (5). Translation of the intein tags utilizes the translation initiation signal (Shine Dalgarno sequence) from the strongly expressed T7 gene 10 protein (φ10).
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(qǐng)下載：
ZK336pTWIN1大腸表達(dá)質(zhì)粒.txt
