摘要
目的对SEN病毒(SENV)开放阅读框(ORF)2基因进行序列测定分析和原核表达,并对表达蛋白进行抗原性分析。方法用聚合酶链反应(PCR)方法扩增SENVORF2基因,对该基因进行序列测定、系统进化分析,双酶切扩增产物与原核表达载体pRSETB构建成重组质粒,诱导表达后进行SDSPAGE和免疫印迹分析。结果该株SENVORF2与北京株(AY072045)核苷酸同源性为97%,氨基酸同源性为59%;与日本株(AB059352)同源性分别为90%和56%。含有SENVORF2质粒的菌株表达相对分子质量约为19×103的融合蛋白,免疫印迹证明该融合蛋白能与SENVDNA阳性血清发生特异反应。结论表达了158个氨基酸的SENVORF2原核表达蛋白具有一定的抗原活性。
Objective To perform phylogenetic and antigenic analysis of open reading frame-2 （OFR2） from SEN virus expressed in E. coli. Methods SEN V OFR2 polymerase chain reaction product was sequenced and phylogneticaUy analyzed, then was expressed by the pRSET B which was transformed into E. coli and induced by IPTG. The fusion protein was detected by Western Blot. Results The percent age of homology of nucleotede sequence and amino acid sequence between Bejing isolate（AY072045） was 97% and 59%, between Japan isolate （AB059352） were 90% and 56% . A 19 ×10^3 fusion protein produced in E. coli. was verified to react with SEN V DNA positive serum by Western Blot. Conclusion Karyota expression protein containing 158 amino acids have definite antigenicity.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2006年第4期352-354,共3页
Chinese Journal of Experimental and Clinical Virology
基金
湖北省科委资助项目(No.2002AA301C32)
关键词
输血传播病毒
基因表达
聚合酶链反应
Transfusion-transmited virus
Gene expression
Polymerase chain reaction
