中国兽医科学2013，Vol.43Issue(1)：65-69,5.
猪附红细胞体DnaJ基因的克隆及真核表达
Cloning and eukaryotic expression of DnaJ gene of Mycoplasma suis
摘要
Abstract
In order to study the function of DnaJ gene of Mycoplasma suis; ,DNAStar,DNAMAN software and ExPASy network server were applied to screen a candidate protein gene DnaJ. A pair of specific primers was designed based on the DnaJ gene in the complete genome sequence of M. Suis in GenBank(NC _015153. 1). The DnaJ gene was amplified by PCR, cloned into the pMD18-T vector and then subcloned into the eukaryotic expression vector pVAX1 to construct the recombinant eukaryotic expression plasmid pVAX-DnaJ. pVAX-DnaJ was transfected into Vero cells and the expression of DnaJ gene was detected by using RT-PCR and indirect fluorescent antibody test(IFAT). The results showed that the length of DnaJ gene is 876 bp, encoding 292 amino acids. The amplified DnaJ gene shared 99% nucleotide identity with that in GenBank. RT-PCR and IFAT results indicated that the DnaJ gene was transiently expressed in Vero cells. This study laid foundations for the research on the biological characteristics of the DnaJ gene and the development of DNA vaccine against M. Suis.关键词
猪附红细胞体/DnaJ基因/真核表达Key words
Mycoplasma suis /DnaJ gene/eukaryotic expression分类
农业科技引用本文复制引用
张影,钱年超,贾立军,薛书江,张守发..猪附红细胞体DnaJ基因的克隆及真核表达[J].中国兽医科学,2013,43(1):65-69,5.基金项目
公益性行业(农业)科研专项(200903036-13) （农业）
吉林省自然科学基金项目(201115230) （201115230）
