摘要
目的构建真核高效表达该基因的载体，为进一步探讨运用高效表达Ｅ１Ａ的重组质粒进行肿瘤基因治疗及研究Ｅ１Ａ基因的作用机理和与其他基因的相互作用提供方法。方法运用分子克隆技术构建２套用较强启动子驱动的表达Ｅ１Ａ的真核重组质粒，并用多种限制性内切酶和ＤＮＡ测序对重组质粒进行鉴定。结果构建的重组质粒分别含有１．３４ｋｂ和１．７７ｋｂＥ１Ａ片段，分别命名为ｐＥ１Ａ１和ｐＥ１Ａ２，且插入的方向均为正向，无重排等异常现象，核苷酸序列无误。结论为人腺病毒５型（Ａｄ５）Ｅ１Ａ基因实验性抗肿瘤基因治疗包括放射增敏研究提供方法学及资料。
Objective Adenovirus type 5 E1A gene has been found to be a tumor
suppressor gene recently. The E1A products are positive and negative regulators for many
other genes which are important in carcinogenesis and tumor metastasis. E1A possesses
activities which suppress transformation, tumorigenesis and malignant progression of tumor
cells. Methods To study the structure, function and antitumor effect of E1A gene, we have
cloned separately a fulllength E1A DNA and another variant lacking the nuclear localization
domain into pcDNA3 vector. The recombinants were cleaved with appropriate endonucleases
and sequenced. Results The orientation of the insert was found to be correct, and no
rearrangement was observed. Conclusions Our work provides data and material for the future
research on the antitumorigenic effects of the E1A gene.
出处
《中华放射肿瘤学杂志》
CSCD
北大核心
1999年第2期106-108,共3页
Chinese Journal of Radiation Oncology
基金
国家自然科学基金
