■ 基本信息
啟動子: | T7,T3 |
復(fù)制子: | pUC |
終止子: | T1 |
質(zhì)粒分類: | 廣宿主系列，枯草桿菌載體 |
質(zhì)粒大小: | 6594bp |
原核抗性: | 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'測序引物: | T7:TAATACGACTCACTATAGGG |
3'測序引物: | 根據(jù)序列設(shè)計(jì)引物 |
■ 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: |
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原核抗性: | Chl |
真核抗性: |
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熒光標(biāo)記: |
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■ 質(zhì)粒簡介
A genomic library of Bacillus lyticus was constructed in lambda GEM 11 vector and screened for the xylanase gene using Congo red plate assay. A 16-kb fragment containing the xylanase gene was obtained which was further subcloned using Mbo I partial digestion in an E. coli pUC 19 vector. A 1.3-kb sub-fragment was obtained which coded for a xylanase gene of Mr 23,650 Da. This fragment was sequenced and the homology was checked with known xylanases. The maximum homology was 97%, which was obtained with an endo xylanase gene from Bacillus species at the DNA level, while the translated sequence showed only one amino acid change from alanine to serine at position number 102. Expression was checked in E. coli, using the native promoter, and an extracellular activity of 5.25 U/mL was obtained. Cloning of the gene was done in Bacillus subtilis using a shuttle vector pHB 201, which resulted in increasing the basal level xylanase activity from 14.02 to 22.01 U/mL.
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列可參考（具體以測序?yàn)闇?zhǔn)）下載：
ZK1022pHB201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。
