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目的构建缺氧诱导因子-1α靶向siRNA重组表达载体。方法根据GeneBank提供的人缺氧诱导因子-1α的核苷酸序列选择设计4对发夹状结构的核苷酸序列,克隆到质粒PGenesil-1中,转化至DH5α菌株,提取质粒进行酶切鉴定和测序分析。结果经基因测序等证实缺氧诱导因子-1α靶向siR-NA重组表达载体碱基序列与设计完全一致。结论缺氧诱导因子-1α靶向siRNA重组表达载体构建成功,为进一步研究肿瘤生物学行为奠定了实验基础。
Objective To construct hypoxia inducible factor-1α target siRNA recombinant expression vector. Methods Based on the nucleotide sequence of human hypoxia inducible factor-1α gene provided by GeneBank, four pairs of hairpin-like nucleotide sequences were designed and cloned into plasmid PGenesil-1 and transformed into DH5α. The plasmid was digested with restriction endonuclease And sequencing analysis. Results The nucleotide sequence of the siR-NA recombinant expression vector targeting hypoxia-inducible factor-1α was completely consistent with the design by gene sequencing. Conclusion The recombinant expression vector of hypoxia-inducible factor-1α targeting siRNA was successfully constructed, which laid the experimental foundation for further study of tumor biological behavior.