犬细小病毒VP2基因真核表达质粒的构建与瞬时表达  

Construction and Transient Expression of Eukaryotic Expression Plasmid for VP2 Gene of Canine Parvovirus(CPV)

作　　者：简子健[1] 马素贞[1] 陈胜男[1] 翟少华[1] 赵森[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业科学》2011年第4期719-723,共5页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区高技术项目(200611107)

摘　　要：【目的】构建犬细小病毒(CPV)VP2基因真核表达质粒,为研究核酸疫苗奠定基础。【方法】根据CPVVP2基因序列设计特殊引物,采用PCR方法从疑似"犬细小病毒"的患犬粪便基因组中扩增VP2基因,将其克隆至真核表达载体pcDNA3.1(+),测序验证后,小白鼠尾静脉注射瞬时表达VP2基因,8 h后取小白鼠肝脏提取总RNA,进行RT-PCR扩增。【结果】在病犬粪便基因组中扩增得到1 755 bp的VP2基因片段,并构建了真核表达质粒pcDNA3.1(+)-VP2,从瞬时表达的小白鼠肝脏总RNA中可扩增到目的条带。【结论】成功构建了犬细小病毒VP2基因真核表达重组质粒,并在小白鼠体内进行了瞬时表达。【Objiective】In order to set the base for studying vaccinum of nucleic acid,VP2 gene eukaryotic expression plasmid of canine parvovirus was constructed in this paper.【Method】The special primer was designed according to the VP2 gene sequence of CPV published in GenBank.VP2 gene was amplified from the stool samples in clinically infected dogs with CPV by PCR.The PCR product was cloned into eukaryotic expression vector pcDNA3.1（＋） to construct a eukaryotic expression plasmid pcDNA3.1-VP2.After sequencing and identifying,the pcDNA3.1-VP2 was injected into mice in vena caudal to induce the transient expression of VP2.The total RNA was extracted from murine livers at 8h after injection and the target strap was obtained by RT-PCR from the total RNA.【Result】1 755 bp of VP2 gene fragment was obtained from the stool samples in clinically infected dogs by PCR.The eukaryotic expression Plasmid,pcDNA3.1-VP2 was constracted and 1 755 bp of a bright stripe was found from total RNA of murine livers by RT-PCR.【Conclusion】The eukaryotic expression plasmid,pcDNA3.1-VP2 was successfully constructed and expressed in murine bodies.

关 键 词：犬细小病毒 VP2基因 真核表达载体 瞬时表达 

分 类 号：S858.292[农业科学—临床兽医学] S188.2[农业科学—兽医学]