利用RT-PCR技术鉴定人ZP3嵌合肽基因在昆虫细胞中的表达

EXPRESSION OF hZP3 CHIMERIC PEPTIDE GENE IN INSECT CELL DETECTED BY RT-PCR

目的 :探讨在转录水平上对人卵透明带蛋白 3(hZP3)嵌合肽在昆虫细胞中的表达鉴定 .方法 :用含目的基因的重组病毒感染昆虫细胞 ,用LiCl法提取RNA ,以其为模板 ,利用一步法RT_PCR反转录出目的DNA .结果 :琼脂糖凝胶电泳显示其PCR产物只有一条DNA条带且与目的基因大小一致 ,而阴性对照未见任何条带 .结论 :hZP3嵌和肽基因在重组病毒感染的昆虫细胞中成功转录 。

Aim:to identify the expression of human ZP3 chimeric peptide(hCP) gene in insect cell at transcriptive level. Methods:Transfected insect cell with recombinant virus containing hCP gene and isolated RNA by LiCl method, and then made it the template for the reverse transcription DNA by one-step RT-PCR. Results: Only one DNA fragment of which the molecular is the same as previously reported in gelose electrophoresis, but not in the control. Conclusion: hCP gene succeeded in transcription in the transfected insect cell with recombinant virus, hence preliminarily proving the expression of target gene.