摘要
根据Genebank中EDSV -AV12 7的序列设计一对引物 ,从EDSV四川株中扩增出六邻体蛋白基因并将其克隆到pUC18中 ,PCR、限制性内切酶分析和序列分析表明 :所扩增、克隆的基因片段包括了EDSV六邻体蛋白基因的完整序列。将克隆的基因正向插入 pBV2 2 0的PRPL 启动子下游的SmaⅠ位点 ,经SDS -PAGE、Western印迹和琼脂扩散试验表明 :六邻体蛋白基因在大肠杆菌中获得了表达并具有免疫学活性。
According to the nucleotide sequence of egg drop syndrome virus AV-127 strain from the Genebank,a pairs oligonucleotide primer was designed and synthesized.With use of polymerase chain reaction (PCR),the hexon protein gene fragment was amplified from the genome of egg drop syndrome virus SG9301 strain that was isolate from Sichuan province in China and the fragment directly inserted into pUC18 vector at EcoRI and PstⅠ site,the recombinant plasmid DNAs were transformed to the competent E.Coli JM109 cells.A positive colony harboring the hexon protein gene was identified by digestion of restriction endonucleases,PCR,sequencing,and the result suggest that the plasmid contain the complete sequence of hexon protein gene.Following,the hexon protein gene was cloned into the expression vector pBV220 and one recominant PPE24 was constructed which highly expressed a 110 KDa protein in E.coli JM109.The result of SDS-PAGE,Western blotting and agar diffusion test indicate the expressed product could react with EDSV postive serum.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2003年第2期172-176,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
四川省青年科技基金 ( 980 0 0 0 2 40 0 75 )
四川省学术带头人培养基金资助 ( 32 0 0 111)。
