■ 基本信息
啟動子: | T7,T3 |
復制子: | pUC |
終止子: | T1 |
質粒分類: | 廣宿主系列，枯草桿菌載體 |
質粒大小: | 6594bp |
原核抗性: | Chl |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度 |
表達宿主: | 枯草芽孢桿菌 |
誘導方式: | IPTG誘導 |
5'測序引物: | T7:TAATACGACTCACTATAGGG |
3'測序引物: | 根據序列設計引物 |
■ 質粒屬性
質粒宿主: | 枯草桿菌 |
質粒用途: | 蛋白表達 |
片段類型: | ORF |
片段物種: |
|
原核抗性: | Chl |
真核抗性: |
|
熒光標記: |
|
■ 質粒簡介
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.
■ 質粒圖譜
■ 質粒序列
質粒序列可參考（具體以測序為準）下載：
ZK1022pHB201枯草胞內質粒.txt
質粒只保證關鍵序列正確，不保證表達效果。
