摘要
用RT-PCR技术对狂犬病病毒(RV)CVS株核蛋白(N)基因进行扩增,经克隆与序列分析,该基因编码450个氨基酸残基。将目的基因克隆到原核表达载体pET28a(+)中,构建了重组质粒pET-N,将其转化表达菌BL21(DE3),于37℃1.0mmol∕L IPTG条件下诱导表达。大肠杆菌菌体裂解产物经SDS-PAGE电泳分析,在分子量约为54kD处出现一新蛋白带,和预期的目的蛋白的分子量相符。质谱鉴定的结果表明成功表达了RV N蛋白,为狂犬病的进一步研究奠定了基础。
N gene of RV CVS strain was amplified by reverse transcriptase-polymerase chain reaction（RT-PCR）. The product was purified and cloned into pMD 18-T,The positive recombinant pMD-N was used for sequencing. The results showed that the complete length of N gene was 1 353bp,which encoded 450 amino acids. The N gene was subcloned into the prokaryotic vector pET28a （＋）. The positive recombinant pET-N was transformed into E, coli BL21 and induced in 1.0mmol/ L IPTG in 37℃ ,SDS-PAGE was performed to analyze the N gene production. Results showed that the protein was highly expressed in E.coli,and the molecular weight was 54kD. The mass spectrum results proved that the N protein of RV CVS strain was successfully expressed,this may provide tools for further study of rabies diseases vaccines and the diagnostic reagent development.
出处
《生物技术通报》
CAS
CSCD
2008年第3期103-106,共4页
Biotechnology Bulletin
基金
甘肃省自然基金项目(3ZS051-A25-076)
甘肃省科技攻关(2GS0S2-A41-006-02)
关键词
狂犬病病毒
核蛋白
原核表达
Rabies virus Nucleoprotein Prokaryotic expression
