摘要
以 Ｐ Ｒ Ｒ Ｓ Ｖ 弱毒株膜蛋白（ Ｍ） 和核衣壳（ Ｎ） 蛋白基因为模板，设计的一对含有 Ｅｃｏ Ｒ Ｉ 和 Ｂａｍ Ｈ Ｉ酶切位点的引物，通过 Ｒ Ｔ Ｐ Ｃ Ｒ 扩增出一约９００ ｂｐ 的 Ｍ Ｎ 基因片段，将此基因片段成功克隆于高效表达载体ｐ Ｂ Ｖ２２０ ，构建成重组质粒ｐ Ｂ Ｖ Ｍ Ｎ，导入大肠杆菌 Ｄ Ｈ５α，经温敏诱导，成功地表达了 Ｍ Ｎ 基因。表达产物经 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 电泳和 Ｗｅｓｔｅｒｎ ｂｌｏｔ 印迹分析，其分子量约３４ ｋ Ｄ，与兔抗 Ｐ Ｒ Ｒ Ｓ Ｖ 高免血清发生反应，经光密度扫描分析，表达产物量占菌体总蛋白的１２ ％ 。该研究为 Ｐ Ｒ Ｒ Ｓ
The primers for RT PCR were designed on the basis of M and N gene sequence of PRRSV. A gene fragment about 900 bp, which has digestion sites of EcoRI and BamHI, was amplified by RT PCR. The M and N gene were cloned into expression vector pBV220 and one recombinant PBVMN was constructed which highly expressed a 34 kD protein in \%E. coli\% DH 5α . The expressed product was identified by SDS PAGE and Western blotting, which occupies 12% of total bacterial protein. The report has laid a basis for the development of molecular diagnostic antigen of PRRSV.
出处
《中国病毒学》
CSCD
1999年第3期244-248,共5页
Virologica Sinica
基金
中华农业科教基金资助项目
关键词
猪
繁殖呼吸综合征
病毒
质粒pV220
表达
M蛋白
Porcine reproductive and respiratory syndrome virus(PRRSV)
pBV220
Expression
M protein
N protein
