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OALib Journal期刊
ISSN: 2333-9721
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新法制备含vegf启动子驱动的cd/tk融合基因重组腺病毒
, PP. 117-120
Keywords: 腺病毒载体,血管内皮细胞生长因子启动子,基因治疗,肿瘤
Abstract:
目的应用简化的细菌内同源重组法高效制备含血管内皮细胞生长因子(vegf)启动子驱动的cd/tk融合基因重组腺病毒。方法先以氯化钙法替代电穿孔法将腺病毒基因组质粒padeasy-1导入bj5183细菌,制备bj5183padeasy-1细菌;再以后者作感受态细菌,与线性化的转移质粒padtrack-vegfp-cdglytk进行同源重组,获得重组腺病毒质粒padeasy-vegfp-cdglytk,转染293细胞,制备含vegf启动子驱动的cd/tk融合基因重组腺病毒。结果构建重组腺病毒质粒的成功率为60%(6/10)。转染293细胞后7~12d可收获病毒。结论简化的细菌内同源重组法简便易行,重组效率较高,易于推广应用。
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