传染性法氏囊病病毒VP_2基因的克隆及其重组鸡痘病毒转移载体的构建  

Cloning of the VP2 Gene of Infectious Bursal Disease Virus and Construction of Its Recombinant Fowlpox Virus Transfer Vector

作　　者：陈忠斌[1] 于康震[1] 王艳华 袁世山[1] 徐宜为[1] 

机构地区：[1]中国农科院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《农业生物技术学报》1996年第3期249-253,共5页Journal of Agricultural Biotechnology

摘　　要：根据已发表的IBDV基因组序列设计并合成一对特异扩增IBDV VP_2cDNA的引物,以纯化的IBDV—D78弱毒疫苗株基因组为模板,利用RT—PCR技术扩增出约1.5kb产物.将PCR产物克隆到pUC19的Smal位点,经酶切图谱分析等方法鉴定出重组VP_2cDNA质粒(pVP_2).将VP_2基因正向插入FPV启动子LP_2EP_2下游,连同痘菌病毒启动子P_(11)启动的LacZ报告基因盒插入FPV转移载体中的FPV复制非必需片段,经鉴定,成功构建了重组FPV—VP_2转移载体.In the study,VP2 gene of infectious bursal disease virus(IBDV)was cloned into pUC19 and arecombinant fowlpox virus (FPV) transfer vector containing the gene was constructed. A pair of 18 -base oligonucleotide primers were designed based on published IBDV genomic sequences,and a 1. 5 kb cDNA fragment was amplified by RT -PCR using genomic RNA of IBDV strain D78 as template,The amplified DNA fragment was inserted into pUC19 at Sma I site. A positive clone harboring IBDV VP2 gene was iden-tide by restriction endonuclease analysis and PCR using the recombinant plasmid DNA as template. Then the VP2 gene was inserted into pSY538 at downstream of LP2EP2 promotor origined from FPV. The LP2EP2 -VP2 cassette and P11 -LacZ cassette were introduced into FPV non-essential fragment in FPV transfer vector pSY681. A recombinanat FPV -VP2 transfer vector pSY781 was identified.

关 键 词：病毒 基因 转移载体 鸡病 IBD 

分 类 号：S858.315.3[农业科学—临床兽医学]