【摘要】 目的:利用基因工程技术对par-4 SAC进行基因克隆并测定其序列,为进一步诱导肿瘤细胞靶向性凋亡的基因治疗奠定基础。方法:采用非对称互补引物/模板法,制备两端含有酶切位点的par-4 SAC的cDNA,PCR产物克隆入pGEM-T Easy载体并转化感受态大肠杆菌E.coli DH5α菌株,随机挑取数个菌落,筛选鉴定并测序。结果:经酶切鉴定、测序分析,表明所插入的基因片断为par-4 SAC基因,与设计完全相同。结论:应用非对称互补引物/模板法克隆par-4SAC基因是成功及可行的。更多还原

【Abstract】 Objective:To use gene engineering technique to clone and sequence the par-4 SAC(prostate apoptosis response-4 gene SAC domain).Methods:By means of asymmetrical primer/template,double stranded cDNA of par-4 SAC was constructed,which had restriction enzymes sites on the two extremes.Then the cDNA was subcloned into the plasmid pGEM-T Easy,which was then transformed into E.coli DH5α and identified by restriction endonuclease digestion and sequence analysis.Results:Evidences of DNA sequence analysis and restriction enzymes digestion showed that the inserted fragment was par-4 SAC cDNA,consistent with the fragment we designed.Conclusion:The par-4 SAC cDNA was cloned successfully.更多还原