应用细菌重组系统构建小鼠Flt3配体的重组腺病毒及其在肝癌细胞中的表达

摘要

Objective　To construct the recombinant adenovirus vector carrying murine Flt3 ligand (AdmFL) and detect the expression of mFL in Hepa1-6 cells following infection by AdmFL. 　 Methods　We amplified mFL from the plasmid and constructed the recombinant adenovirus vector using a new simplified bacterial homologous recombination system, then infected Hepa1-6 cells and detected the expression of mFL by RT-PCR. 　Results We successfully constructed the recombinant adenovirus vector carrying mFL and detected its expression in infected Hepa1-6 cells. 　 Conclusions　The construction of adenovirus vector carrying mFL paves the way for further application in cancer gene therapy.%目的构建携带小鼠Flt3配体（murine flt3 ligand,mFL）的重组腺病毒载体并检测它在感染后的小鼠肝癌细胞中的表达,为肝癌基因治疗提供实验基础。方法 PCR扩增含有mFL的质粒，应用高效细菌内同源重组系统构建携带mFL的重组腺病毒，感染小鼠肝癌细胞株Hepa1-6，一步法提取细胞总RNA，RT-PCR检测mFL的表达。结果构建了携带mFL的重组腺病毒，并在感染后的Hepa1-6细胞中检测到了mFL的mRNA。结论应用该细菌重组系统成功地构建了携带mFL的重组腺病毒载体，为下一步的基因治疗研究奠定了基础。