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目的克隆Vasohibin基因,包装制备成8型重组腺相关病毒,感染肌肉组织,并观察其在肌肉组织中的表达。方法以Vasohibin基因全长cDNA为模板,应用聚合酶链反应(PCR)方法扩增出Vasohibin基因,采用三质粒磷酸钙共转染HEK293细胞,包装制备成8型重组腺相关病毒,感染小鼠(6只)肌肉组织,免疫组织化学方法检测Vasohibin基因的表达,肌肉组织内染有棕色信号者为阳性。结果成功克隆Vasohibin基因,并制备成8型重组腺相关病毒,感染肌肉组织后实验组小鼠肌肉组织切片呈现广泛棕黄色阳性信号,阳性率为100％(6／6),对照组呈阴性(0／6), 未见有阳性表达。结论 8型重组腺相关病毒能够有效介导Vasohibin基因在小鼠肌肉组织中的表达。
Objective To clone Vasohibin gene and package it into recombinant adeno-associated virus type-8 virus to infect muscle tissue and observe its expression in muscle tissue. Methods Vasohibin gene was amplified by polymerase chain reaction (PCR) using the full-length cDNA of Vasohibin as a template. HEK293 cells were co-transfected with the three plasmid calcium phosphate and packaged into recombinant adeno-associated virus type-8 6) muscle tissue. Immunohistochemistry was used to detect the expression of Vasohibin gene. The muscular tissue with brown signal was positive. Results The Vasohibin gene was successfully cloned and the recombinant adeno-associated virus 8 was successfully prepared. Muscle tissue sections of the experimental group showed extensive brown-yellow positive signals after being infected with muscle tissue. The positive rate was 100% (6/6) and negative in the control group 0/6), no positive expression was found. Conclusion Type 8 recombinant adeno-associated virus can effectively mediate the expression of Vasohibin gene in mouse muscle tissue.