Polycistronic expression of CGA-N46 gene in Bacillus subtilis DB1342

Polycistronic expression of CGA-N46 gene in Bacillus subtilis DB1342
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CGA-N46基因在枯草芽孢杆菌DB1342中的多顺反子表达

DOI:
10.5897/ajmr10.796
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发表时间:
2013-06
影响因子:
--
通讯作者:
Qian-cheng Xiong
Qian-cheng Xiong
中科院分区:
--
文献类型:
--
作者:
Rui-fang Li;Bin Wang;Yan-jie Yi;Liang Huang;Qian-cheng Xiong

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CGA-N46是一种小型抗真菌衍生肽,由人嗜铬粒蛋白an端第31 ~ 76个氨基酸组成。将含有核糖体结合位点(RBS)、sacB信号肽核苷酸序列(sacB s)、CGA-N46编码序列(CGA-N46)和终止密码子的单拷贝、双拷贝和3拷贝外源基因片段串联构建到pSBPTQ的多克隆位点上。将CGA-N46基因单顺子、双顺子和三顺子表达载体转染枯草芽孢杆菌DB1342。工程菌株DB1342(p-N46)、DB1342(p-2N46)和DB1342(p-3N46)的表达结果表明,单顺反和双顺反表达系统产生了少量的CGA-N46,而三顺反表达系统提高了肽的产生。本研究结果有助于提高工程菌株小肽的产量。
CGA-N46 is a small antifungal derived peptide and consists of the 31st to 76th amino acids of the N-terminus of human Chromogranin A. Polycistronic expression of CGA-N46 gene in Bacillus subtilis DB1342 was used to improve its production. Single, double and three copies of exogenous gene fragment which contained ribosome binding site (RBS),sacB signal peptide nucleotide sequence (sacB s), CGA-N46 encoding sequence (cga-N46) and stop codon were tandem constructed into the muticloning sites of pSBPTQ. The engineered expression plasmids which bore monocistronic, bicistronic and tricistronic expression cassettes of the CGA-N46 gene were transformed into the competent B. subtilis strain DB1342. The expression results of the engineered strains DB1342(p-N46), DB1342(p-2N46) and DB1342(p-3N46) demonstrated that monocistronic and bicistronic expression systems produced low amounts of CGA-N46 while the tricistronic expression system improved the production of the peptide. Our results are helpful for improving the production of small peptides by engineered strains.   Key words: Antifungal peptide, CGA-N46, Polycistronic expression, Secreted expression.
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