摘要
目的:构建白血病相关基因LRP16真核表达质粒,并检测其在人子宫内膜癌HEC-1-B细胞中的表达。方法:从人子宫内膜癌HEC-1-B细胞中提取总RNA,应用PCR技术,扩增获得LRP16基因编码序列片段,克隆入真核表达载体EX-Y2069-M29,对重组质粒进行酶切和测序鉴定后,以脂质体介导法转染至HEC-1-B细胞,采用Western blot法检测LRP16蛋白的表达。结果:酶切和测序结果证明LRP16基因真核表达质粒EX-Y2069-M29的DNA序列完全正确,将其转染HEC-1-B细胞后,LRP16蛋白表达明显增加。结论:LRP16基因重组真核表达质粒EX-Y2069-M29构建成功,并能在HEC-1-B细胞中表达,为进一步研究LRP16基因奠定了基础。
Objective： To construct the eukaryotic expression vectors of leukemia related proteinl6（LRP16） plasmid and de- tect its expression in human endometrial carcinoma HEC-1-B cells. Methods： The coding sequence of LRP16 was generated by PCR using total RNA extracted from the HEC -1-B cells. LRP16 gene was cloned into the eukaryotic expression vector EX-Y2069-M29. After restriction enzyme analysis and sequence identification, the recombinant plasmid was transfected into HEC- 1-B cells with liposome mediated method. The expression of LRP16 was detected by Western blot. Results： The results of enzyme analysis and sequencing both identified DNA sequence of LRP16 gene eukaryotic expression plasmid EX-Y2069-M29 correctly. The expression of LRP16 in- creased obviously after transfection into HEC-1-B cells. Conclusion： LRP16 gene eukaryotic expression plasmid EX-Y2069-M29 was constructed successfully and it can be expressed in HEC-1-B ceils. This provides experimental basis for further study on the function of LRPI6.
出处
《肿瘤预防与治疗》
2013年第3期117-121,共5页
Journal of Cancer Control And Treatment
基金
四川省泸州医学院课题(07055FF09)
