纹皮蝇HypoderminC基因真核表达质粒构建及小鼠免疫试验  

Construction of Hypoderma lineatum Hypodermin C Gene Eukaryotic Expression Plasmid and Immune Tests in Mice

作　　者：白红岩[1] 刘春霞[2] 呼和巴特尔[1] 王文龙[2] 

机构地区：[1]内蒙古农业大学兽医学院农业部动物临床诊疗技术重点实验室,内蒙古呼和浩特010018 [2]内蒙古农业大学生命科学学院,内蒙古呼和浩特010018

出　　处：《动物医学进展》2012年第8期1-5,共5页Progress In Veterinary Medicine

基　　金：内蒙古自然科学基金(2009BS0403);内蒙古农业大学博士科研启动金(BJ07-48)

摘　　要：根据GenBank发表的Hypodermin C(HC)基因序列设计引物,以原核表达载体pET-HC为模板,PCR扩增HC基因,将克隆基因插入到真核表达载体pVAX1中,构建重组真核表达质粒pVAX1-HC。将重组质粒pVAX1-HC转染小鼠胎儿成纤维细胞,以间接免疫荧光法检测HC基因在细胞中表达。用真核表达质粒pVAX1-HC经双侧胫前肌注射昆明小鼠,对小鼠进行体液免疫和细胞免疫研究。结果表明,小鼠血清抗体水平和淋巴细胞增殖水平明显高于非免疫对照组。本研究成功构建了真核表达质粒pVAX1-HC,为纹皮蝇核酸疫苗研发奠定了基础。The primers were designed according to sequence of Hypodermin C （HC） gene in GenBank, and HC gene was amplified by PCR using the pET-HC as template, then cloned into the eukaryotic expression vector pVAX1 to construct a recombinant expression plasmid named pVAX1-HC. The recombinant plas- mid pVAX1-HC was transfected into the mice embryonic fibroblasts （MEF）, and Hypodermin C expres- sion was detected in vitro by indirect immunofluorescence method. After intramuscular injection into Kun- ming mice with pVAX1-HC in order to research the humoral immunity and cellular immunity, the results showed that the serum antibody levels and lymphocyte proliferation levels were significantly higher than that in the control group. This eukaryotic expression plasmid can develop as novel DNA vaccine of hypo- dermosis.

关 键 词：纹皮蝇 HYPODERMIN C基因 真核表达质粒 免疫 小鼠 

分 类 号：Q786[生物学—分子生物学] S855.9[农业科学—临床兽医学]