[1]黎小军,林陈水.Burkholderia cepacia XYU-6脂肪酶的克隆及其细胞表面展示[J].江西师范大学学报(自然科学版),2015,(05):502-506.
 LI Xiaojun,LIN Chenshui.The Cloning and Cell Surface Display of a Lipase from Burkholderia cepacia XYU-6[J].,2015,(05):502-506.
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Burkholderia cepacia XYU-6脂肪酶的克隆及其细胞表面展示()
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《江西师范大学学报》(自然科学版)[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
502-506
栏目:
出版日期:
2015-10-01

文章信息/Info

Title:
The Cloning and Cell Surface Display of a Lipase from Burkholderia cepacia XYU-6
作者:
黎小军;林陈水
新余学院医学与生命科学学院,江西新余,338000;浙江工业大学药学院,浙江杭州,310032
Author(s):
LI Xiaojun;LIN Chenshui
关键词:
脂肪酶克隆细胞表面展示表达
Keywords:
lipasecloningcell surface displayexpression
分类号:
Q812
文献标志码:
A
摘要:
通过分析GenBank中Burkholderia cepacia脂肪酶的序列,设计简并引物,采用同源克隆的策略,成功地从B. cepacia XYU-6菌株中克隆到脂肪酶基因bcl,其大小为1095 bp,编码364个氨基酸( GenBank登陆号KR233260).将bcl基因与质粒pET-28b(+)连接并转化大肠杆菌,使脂肪酶BCL的大肠杆菌胞内过表达,其活力是野生菌的12.9倍.通过将bcl基因克隆到细胞表面展示载体pZXL中,构建脂肪酶BCL的细胞表面展示工程菌,使BCL在Lpp-OmpA引导下定位于大肠杆菌细胞表面,其活力是野生菌的3.9倍.研究结果为脂肪酶BCL后续的分子改造和应用奠定基础.
Abstract:
The lipase gene bcl was isolated from Burkholderia cepacia XYU-6 by homologous cloning method using degenerate primers,which was design based on the analyzing the sequences of lipases from B. cepacia. The gene bcl contained a 1 095 bp open reading frame encoding a protein of 364 amino acids( GenBank accession number:KR233260 ). The gene bcl was cloned into the expression vector pET-28 b( +),and intracellular overexpressed in biologically active in Escherichia coli. Meanwhile,the gene bcl was cloned into the cell-surface display vector pZXL and transformed into E. coli. The lipase BCL was successfully achieved on the cell surface of engineering strain using the anchoring motif Lpp-OmpA. The two recombinant strains demonstrated 12. 9-fold and 3. 9-fold of lipase activity compared to the wild B. cepacia XYU-6,respectively. This study paves the way for the further research of the lipase for protein engineering and application in the industry.

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备注/Memo

备注/Memo:
江西省青年科学基金(20122BAB214011)
更新日期/Last Update: 1900-01-01