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目的构建携带人次级淋巴组织趋化因子的重组腺病毒载体(Ad-6Ckine),并观察其在真核细胞中的表达,为进一步的肿瘤基因治疗研究奠定基础。方法 RT-PCR法从人淋巴结中扩增出人次级淋巴组织趋化因子cDNA,利用AdMax腺病毒包装系统在细胞内同源重组出Ad-6Ckine,然后用Ad-6Ckine感染真核细胞,观察6Ckine蛋白在真核细胞中的表达。结果成功构建了重组Ad-6Ckine,PCR法鉴定无野生型腺病毒的污染,构建的Ad-6Ckine所携带的6Ckine基因能在真核细胞中得到表达。结论 AdMax腺病毒包装系统是一种简便高效的重组腺病毒构建方法,构建的重组Ad-6Ckine可用于肿瘤基因治疗的体内外研究。
Objective To construct a recombinant adenovirus carrying human secondary lymphoid chemokine (Ad-6Ckine) and to observe its expression in eukaryotic cells, which will lay the foundation for further research on gene therapy of tumors. Methods Human secondary lymphoid tissue chemotaxis cDNA was amplified by RT-PCR from human lymph nodes. Ad-6Ckine was homologously recombined into AdMax adenovirus packaging system and then infected with eukaryotic cells using Ad-6Ckine Expression of 6Ckine protein in eukaryotic cells. Results Recombinant Ad-6Ckine was successfully constructed. The contamination of wild-type adenovirus was identified by PCR. The constructed 6Ckine gene of Ad-6Ckine was expressed in eukaryotic cells. Conclusion The AdMax adenovirus packaging system is a simple and efficient method for constructing recombinant adenovirus. Recombinant Ad-6Ckine can be used in vitro and in vivo for gene therapy of tumors.