小熊猫源犬瘟热病毒DNA疫苗构建及其在真核细胞的表达  

Construction of DNA Vaccine Containing H and F Gene of Canine Distemper Virus From Red Panda and its Expression in Eukaryotic Cells

作　　者：张晓明[1,2] 陈武[1] 彭仕明[1] 

机构地区：[1]广州动物园,广州510070 [2]广州海洋馆,广州510070

出　　处：《野生动物学报》2015年第3期279-283,共5页CHINESE JOURNAL OF WILDLIFE

摘　　要：根据Gen Bank中发表的犬瘟热病毒(CDV)的基因组序列设计了H和F基因的2对引物,通过RT-PCR扩增出从发病小熊猫分离的CDV的H和F基因,并将其克隆入到18T-simple载体上进行测序。使用Kpn I和Xho I限制性内切酶对重组载体进行双酶切并将目的基因克隆到pcDNA 3.1表达载体上,构建完成后转染293T细胞,转染48 h后,再利用RT-PCR和Western-blot检测目的基因的转录和目的蛋白的表达。结果表明,表达载体经转染后,在293T细胞中,目的基因均能得到转录而且正确表达。这为下一步基因疫苗的研究奠定了良好的基础。The primers of H and F genes of the CDV were designed according to sequences of the CDV from GeneBank and used for amplifying of H and F genes of CDV from red panda by RTPCR.Two recombinant plasmids were constructed with 18T-simple vector for cloning and sequencing after insertion of the H and F genes.Two expressing vector plasmids were constructed with pcDNA 3.1 vector in the same way,and transfected into the 293 T cell.Transcriptions of targeted genes were detected by RT-PCR and the expressing target proteins were detected by the WesternBlot method.The transcription of objective genes and expression of interesting proteins could be detected at 48 h post-transfection.This will lay a foundation for the research of DNA vaccine of CDV.

关 键 词：犬瘟热病毒 DNA疫苗 载体构建 真核表达 

分 类 号：S858.93[农业科学—临床兽医学]