%0 Journal Article
%T Efficient fusion expression of G13 domain derived from granulysin in Escherichia coli<br>颗粒裂解肽G13结构域在大肠杆菌中的高效融合表达
%A Xiaoqiang Liu
%A Xiangdong Zh
%A Yazhong Xiao
%A Jinhuan Yang
%A Nengshu Li
%A <br>刘小强
%A 查向东
%A 肖亚中
%A 杨金环
%A 李能树
%J 生物工程学报
%D 2009
%I 
%X The G13 domain derived from granulysin shows high antimicrobial activities against Gram-positive and Gram-negative bacteria but does not lyse Jurkat cells or liposomes. To explore a new approach for high expression of the G13 domain, we fused the sequence encoding G13 to thioredoxin (Trx) gene to construct the recombinant expression vector (pThioHisA-G13). A cyanogen bromide (CNBr) cleavage site was introduced between the Trx and G13 to facilitate final release of the recombinant G13. The recombinant expression vector, pThioHisA-G13, was transformed into E. coli BL21 (DE3). Upon induction by IPTG, Trx-G13 fusion protein was expressed and took the form of inclusion bodies counting 58% (W/W) of total cellular proteins. The inclusion body was solved by urea (8 mol/L) and then cleaved by CNBr. We purified the recombinant peptide G13 by one-step cation exchange chromatography. Results of agarose diffuse assay analysis indicated that the recombinant G13 exhibited antibacterial activity. The procedure described in this study will provide a reliable and simple method for highly efficient production of some cationic antimicrobial peptides.
%K Granulysin
%K G13 domain
%K fusion expression
%K cationic antimicrobial peptide<br>颗粒裂解肽
%K G13结构域
%K 融合表达
%K 阳离子抗菌肽
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=283461636DCBBAE17CEAAB8345E04ABF&yid=DE12191FBD62783C&vid=C5154311167311FE&iid=0B39A22176CE99FB&sid=6CCE24D86D03D083&eid=6A73B36E85DB0CE9&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=18