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目的:构建并鉴定含14-3-3蛋白抑制肽R18的重组腺病毒,为研究14-3-3蛋白的功能提供基础工具。方法:用同源重组方法构建含14-3-3蛋白抑制肽R18的复制缺陷型腺病毒载体(AdR18),并加以鉴定、扩增,以获得高滴度AdR18病毒液,体外感染乳大鼠心肌细胞,检测目的基因表达。结果:将构建的重组腺病毒载体AdR18感染乳大鼠心肌细胞并表达48h后,蛋白印迹结果显示AdR18感染组有明显R18的表达,对照组无表达。结论:腺病毒载体可高效率导入外源基因在心肌细胞中高表达。
OBJECTIVE: To construct and identify a recombinant adenovirus containing the 14-3-3 protein inhibitor R18, which provides a basic tool for studying the function of 14-3-3 protein. METHODS: The replication-deficient adenovirus vector (AdR18) containing the 14-3-3 protein inhibitor R18 was constructed by homologous recombination and identified and amplified to obtain high titer AdR18 virus solution. Cardiomyocytes, detect the expression of the target gene. Results: The constructed recombinant adenovirus AdR18 infected rat myocardial cells and expression of 48h, Western blot results showed that AdR18 infection significantly R18 expression, the control group no expression. Conclusion: Adenovirus vectors can efficiently introduce foreign genes into cardiomyocytes.