摘要： 以山西省农业科学院畜牧兽医研究所预防兽医研究室克隆的pG-F和pG-HN质粒为模板,PCR扩增麻鸡新城疫病毒SX-1株F基因和HN基因,并以由15个柔性氨基酸构成的柔性短肽作为Linker将F及HN基因片段体外连接,插人pCI-neo真核表达载体,构建麻鸡新城疫病毒SX-1株F基因和HN基因共表达载体pCI-F-HN,并在Vero细胞中转染表达,进行间接免疫荧光检测.结果显示,麻鸡新城疫病毒SX-1株F-HN组合基因在Vero细胞中得到较高水平的表达,在荧光显微镜下能够观察到较强的荧光,说明F-HN组合蛋白具有良好的反应原性.为下一步研究高效亚单位基因工程苗奠定基础.

关键词: 麻鸡新城疫病毒, SX-1株, F-HN基因, 共表达

Abstract: Based on The pG-F and pG-HN cloning plasmid templated in our lab,Newcastle disease virus strain SX-1 sequences of F gene and HN gene isolated from?spotted chicken were PCR amplified,15 amino acids as flexible short peptide Linker connected F and HN genetic fragments in vitro was inserted into pCI-neo eukaryotic expression vector to construct the co-expression vector pCI-F-HN of Newcastle disease virus strain SX-1 sequences of F gene and HN gene and transfected in Vero cells,and carried on expression of indirect immune fluorescence detection.The results showed that virus SX-1 strain F-HN combination of genes in Vero cells had high level expression and in fluorescence microscopy strong fluorescence signal could be observed,which suggested that F-HN combination protein had good reactionogenicity.It provids for further research efficient subunits genetic engineering?vaccine in foundation.

Key words: Spotted chicken newcastle disease virus, Strain SX-1 sequences, F-HN gene, Co-expression

中图分类号: 

• S831

• Q782