■ 基本信息
別名: | pHT-01 |
啟動(dòng)子: | Pgrac |
復(fù)制子: | pUC |
質(zhì)粒分類: | 廣宿主系列，枯草桿菌載體 |
質(zhì)粒大小: | 7956bp |
原核抗性: | Amp |
真核抗性: | Chl |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度 |
表達(dá)宿主: | 枯草芽孢桿菌 |
誘導(dǎo)方式: | IPTG誘導(dǎo) |
5'測(cè)序引物: | 根據(jù)序列設(shè)計(jì)引物 |
3'測(cè)序引物: | 根據(jù)序列設(shè)計(jì)引物 |
備注: | 枯草桿菌IPTG誘導(dǎo)胞內(nèi)表達(dá)空載體 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 枯草桿菌 |
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: |
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■ 質(zhì)粒簡(jiǎn)介
All vectors use the strong promoter preceding the groESL operon of Bacillus subtilis fused to the lac operator allowing their induction by addition of IPTG. While the background level of expression of these expression cassettes is very low in the absence of the inducer, an induction factor of about 1,300 was measured using the bgaB reporter gene. The amount of recombinant protein produced after addition of IPTG may represent 10 and 13%, respectively, of the total cellular protein. High level secretion of amyQ α-amylase and cellulase A and B of Clostridium thermocellum was demonstrated. An efficient Shine-Dalgarno sequence as well as a multiple cloning site (BamH I, Xba I, AatII, SmaI) were also inserted. To obtain secretion of recombinant proteins, the coding region for the signal peptide of the amyQ gene encoding an α-amylase was fused to the SD sequence of pHT01, thereby constructing pHT43.
大腸桿菌-枯草芽孢桿菌穿梭質(zhì)粒的表達(dá)載體pHT01可以在枯草芽孢桿菌中高效表達(dá)重組外源蛋白。載體基于強(qiáng)σA-依賴性啟動(dòng)子的枯草桿菌groE操縱子，通過添加lac操縱子改造成為一種高效可控的（IPTG誘導(dǎo)的）啟動(dòng)子。
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK1011pHT01枯草胞內(nèi)質(zhì)粒.txt
質(zhì)粒只保證關(guān)鍵序列正確，不保證表達(dá)效果。
