卵形巴贝斯虫AMA1基因的克隆与原核表达  

Prokaryotic expression of AMA1 gene from Babesia ovata

作　　者：田万年 刘杰 张馨月 薛书江[2] 贾立军[2] 张守发[2] TIAN Wan-nian;LIU Jie;ZHANG Xin-yue;XUE Shu-Jiang;JIA Li-jun;ZHANG Shou-fa(College of Animal Science,Jilin Agricultural Science and Technology College,Jilin 132101,China;College of Agriculture,Yanbian University,Yanji 133002,China)

机构地区：[1]吉林农业科技学院动物科技学院,吉林吉林132101 [2]延边大学农学院,吉林延吉133002

出　　处：《中国兽医杂志》2019年第7期41-43,47,I0001,共5页Chinese Journal of Veterinary Medicine

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20190981KJ)

摘　　要：为了解卵形巴贝斯虫AMA1基因蛋白特性及免疫活性,本试验用卵形巴贝斯虫特异性引物进行PCR扩增,克隆到pGEX-4T-1中构建BoAMA1重组pGEX-4T-1表达载体,经IPTG诱导表达后,进行SDS-PAGE、Western Blot分析。结果显示,克隆的基因片段长1042 bp,编码347个氨基酸,与GenBank中相应基因序列(KT312793)的同源性为99.8%。SDS-PAGE分析和Western Blot分析表明,BoAMA1蛋白分子质量约为68 ku,具有较好反应原性。本试验为卵形巴贝斯AMA1基因疫苗研究奠定了基础。In order to understanded the characteristics of Babesia ovata AMA1protein and analyze its immunogenicity,the gene AMA1 of Babesia ovata was amplified by PCR.A prokaryotic expression plasmid was constructed by inserting the BoAMA1gene into pGEX-4T-1.The expression was induced by IPTG,and the expressed protein was analyzed by SDS-PAGE and Western Blot.Results showed that the amplified fagment was 1042 bp,encoded 347amino acids,and shared 99.8%homology with the corresponding gene sequence in GenBank(KT312793).SDS-PAGE analysis showed that the recombinant protein was expressed in E.coli with 68 ku in molecular mass.Western Blot analysis indicated that BoAMA1protein possessed good reactinogenicity.These results will facilitate future study of the Babesia ovata.

关 键 词：卵形巴贝斯虫 AMA1基因 原核表达 

分 类 号：R852.7[医药卫生—航空、航天与航海医学]