融合头的改变对抑制重组抗菌肽毒性的影响(英文)  

The effects of fusion partners on the inhibition of the toxicity of recombinant antimicrobial peptides to expression host

作　　者：王作利[1] 查向东[1] 周鹏[1] 刘小强[1] 杨金环[1] 方红[1] 

机构地区：[1]安徽大学生命科学学院安徽省生态工程与技术重点实验室,合肥230039

出　　处：《中国抗生素杂志》2010年第2期91-95,137,共6页Chinese Journal of Antibiotics

基　　金：安徽省高校省级自然科学研究重点项目:抗菌肽高效重组表达与结构功能关系研究(KJ2010A025)

摘　　要：为进一步完善G13结构域的原核表达系统,人工合成编码G13的基因片段,PCR扩增后克隆于原核表达载体pET28a中,构建的重组质粒pET28a-G13转化于E.coli BL21(DE3)中。另外对pET28a-G13进行突变,改变其G13片段N端上游处的融合头的电荷数,转化于E.coli BL21(DE3)中。构建pThiohisA突变体并与pET28a-G13共转化于E.coli BL21(DE3)中。用诱导剂诱导所构建的各个工程菌,然后比较A600值变化及蛋白表达量,分析不同的融合头对G13毒性抑制效果,发现融合头对G13毒性抑制效果与其净负电荷数及酸性氨基酸的相对位置有关。In order to improve prokaryotic expression system for the G13 domain of granulysin, the DNA sequence encoding G13 domain was synthesized, amplified by PCR and cloned into the expression vector pET28a, and transformed to E.coli BL21（DE3）. In addition, we mutated pET28a-G13 to change the charge of the fusion partner. A mutated pThioHisA was also constructed and co-transformed with pET28a-G13 into E.coli BL21 （DE3）. After induction with the inducers, the A600 values and protein expression levels were compared to analyze the effects of different fusion partners on the inhibition of the toxicity of G 13. It was found that the inhibition effect of the fusion partners was greatly affected by the net negative charge and the relative location of the acidic amino acid residues.

关 键 词：阳离子抗菌肽 融合头 共表达 重组表达 

分 类 号：Q786[生物学—分子生物学]