Hengjun Gao, Hongyin Zhu, Weiqi Gu, Y. Lou, W. Ren, SH Xiao
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引用次数: 0
摘要
目的:利用白细胞介素-2 (IL-2)进行肿瘤基因治疗已引起人们极大的兴趣,因为这种细胞因子具有强大的抗肿瘤作用。目前正在研究一种利用痘苗病毒载体的有前途的癌症基因治疗新方法。本研究的目的是构建牛痘真核表达载体pMJ601,该载体含有人IL-2 (IL-2;pMJ601hIL-2),可用于胃癌的治疗。方法:采用质粒提取、琼脂糖凝胶电泳、酶切分析、连接、制备感态细胞、转化、DNA序列分析等基因工程技术,将hIL-2基因克隆到pBluescript II SK+/ -和pMJ601载体中,并进行鉴定。结果:从EcoR1-BamHI酶切酶中提取的pLXSN hIL-2 DNA片段成功克隆到pBluescript II SK+/ -中。此外,pBluescript II SK+/ -的hIL-2 DNA也通过salii - bamhi酶切成功克隆到pMJ601中。结论:构建表达hIL-2基因的pMJ601载体是实现胃癌基因治疗的重要一步。
Construction and identification of the eukaryotic expression vector of vaccinia virus expressing human interleukin-2
OBJECTIVE: Cancer gene therapy using interleukin-2 (IL-2) has generated much interest because of the potent antitumor effect of this cytokine. There is ongoing research into one promising new gene therapy approach for cancer using the vaccinia virus vector. The purpose of this study was to construct a vaccinia eukaryotic expression vector, pMJ601, which contains human IL-2 (hIL-2; pMJ601hIL-2) and can be used for the treatment of gastric carcinomas.
METHODS: Genetic engineering techniques such as plasmid extraction, agarose gel electrophoresis, restriction analysis, ligation, preparation of competent cells, transformation and DNA sequence analysis were used to clone the hIL-2 gene into pBluescript II SK+/– and pMJ601 and identify these vectors.
RESULTS: Fragments of hIL-2 DNA from pLXSN from an EcoR1–BamHI restriction enzyme digest were successfully cloned into pBluescript II SK+/–. In addition, hIL-2 DNA from pBluescript II SK+/– with a SalI-BamHI restriction enzyme digest was also successfully cloned into pMJ601.
CONCLUSION: Constructing a pMJ601 vector that expresses the hIL-2 gene is an important step toward being able to treat gastric carcinoma using gene therapy.