【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 VP₂cDNA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VP₂cDNA质粒（pVP₂）.将VP₂基因正向插入FPV启动子LP₂EP₂下游,连同痘菌病毒启动子P₁₁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—VP₂转移载体.更多还原

【Abstract】 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.更多还原