摘要
用RT PCR方法扩增了新城疫病毒 (NDV)的完整HN基因序列 ;设计了 1对带有EcoRⅠ限制位点的引物 ,用PCR扩增出了HN基因片段。将该片段克隆至 pSOC质粒SOC基因 3′端 ,成功构建了重组质粒pSOC HN。用该重组质粒转化E .coliBL 2 1(DE3)感受态细胞 ,以终浓度 1mmol/mL的IPTG诱导表达。在SDS PAGE凝胶上可检测到分子质量约 6 7ku的特异条带 ,蛋白质印迹法证实 。
HN gene with length of 1 713 bp of Newcastle disease virus was amplified by RT-PCR. The PCR product was purified and cloned into pGEM-T Easy vector and the recombinant plasmid was (sequenced). According to the HN gene sequencing result, another pair of primers containing EcoRⅠrestriction site was designed to amplify the HN fragment coding the globular domain, i.e. the main functional (region) of HN protein. Then the HN fragment was inserted into the 3′ terminal of SOC gene of pSOC plasmid. The recombinant plasmid pSOC-HN was conducted into E.coli BL-21(DE3) to induce HN gene expression with 1 mmol/L IPTG. A SOC-HN fusion protein band with molecular weight of 67 ku was dectected on the SDS-PAGE gel and nitrocellulose membrane,and the expression products can specifically reacted with the antisera against Newcastle disease virus.
出处
《中国兽医科技》
CAS
CSCD
北大核心
2004年第8期27-31,共5页
Chinese Journal of Veterinary Science and Technology
基金
国家高技术研究发展计划 (86 3)项目 (2 0 0 2AA2 4 5 0 5 1)
