Gal-13成熟肽基因在毕赤酵母中的表达与抑菌活性分析  

Expression of chicken β-defensin-13 mature peptide gene in Pichia pastoris and analysis of its antimicrobial activity

作　　者：许国洋 付利芝[2] 杨金龙[2] 曹国文[2] 张素辉[2] 邱进杰[2] 潘康成[1] 

机构地区：[1]四川农业大学动物医学院,四川雅安625000 [2]重庆市畜牧科学院,重庆402460

出　　处：《中国兽医学报》2015年第8期1260-1263,共4页Chinese Journal of Veterinary Science

基　　金："教育部长江学者和创新团队发展计划"创新团队资助项目(IRT0848)

摘　　要：扩增鸡β-防御素-13(Gal-13)成熟肽基因序列,构建酵母重组表达载体pPICZαA-Gal-13,通过SacⅠ限制性内切酶线性化处理后,电击转化毕赤酵母X-33,并对利用甲醇诱导后的重组菌株表达产物进行表达规律分析和抑菌活性研究。结果表明,扩增到的Gal-13成熟肽基因序列为108bp,共编码36个氨基酸;筛选到的重组菌株均为Mut+表型,其诱导表达上清经Tricine-SDS-PAGE凝胶电泳后,发现其在约5 900的位置出现目的条带,抑菌试验发现其对沙门菌(CVCC534)和金黄色葡萄球菌(ATCC25923)具有抑菌活性。In order to analyze the expression principle and the bacteriostatic activity of chickenβ-defensin-13（Gal-13）,the mature peptide gene sequence of Gal-13 was augmented and inserted into pPICZαA to build the recombinant plasmid pPICZαA-Gal-13,and the correct recombinant plasmid was linearized and transformed into Pichia pastoris strain X-33 by electroporation.The result showed that the mature peptide gene of Gal-13 consists of 312 bp,encoding 36 amino acids.The recombination strain,which was induced by methyl alcohol,was Mut＋phenotype,and the target protein secreted into the microbial supernatant showed at a level of 5 900 according to the TricineSDS-PAGE.The antimicrobial bioactivity tests showed that Gal-13 had antimicrobial bioactivity against Salmonella gallinarum（CVCC534）and Staphylococcus aureus（ATCC25923）.

关 键 词：Gal-13 成熟肽 诱导表达 抑菌活性 

分 类 号：S852.4[农业科学—基础兽医学]