【摘要】 目的:构建携带Myc标签的乳腺癌耐药蛋白(BCRP)基因的真核表达载体,获得Myc-BCRP融合表达蛋白,检测其对喉癌细胞Hep-2生长的影响。方法:采用PCR技术从乳腺文库中扩增人BCRP基因,并将其正确插入Myc载体;将重组质粒与空载体分别转染喉癌细胞Hep-2后,Western印迹检测表达情况,并进行生长曲线实验。结果:双酶切和测序鉴定表明Myc-BCRP真核表达质粒构建成功;转染Hep-2细胞后获得表达,生长曲线实验结果表明,BCRP可促进喉癌细胞生长。结论:构建了携带Myc标签的人BCRP基因的真核表达载体,重组载体能在Hep-2细胞中表达,且能促进该细胞的生长。本实验为进一步研究BCRP在喉癌中的功能奠定了基础。更多还原

【Abstract】 Objective: To construct the eukaryotic expression vector of human breast cancer resistance protein(BCRP) labeled with Myc tag and detect its activity. Methods: Human BCRP coding region was amplified from human mammary c DNA library by PCR and cloned into Myc vector. Human Hep-2 cells were transfected with the recombinant plasmid Myc-BCRP and the expression was detected by Western blot. The proliferation in Hep-2 cells was identified by growth curve. Results: BCRP eukaryotic expression vector labeled with Myc was confirmed by double digestion identification. The inserted fragment was identified to be correct by sequencing. Human BCRP protein was expressed in human Hep-2 cells as shown by Western blot. The result demonstrated that BCRP protein could promote proliferation in Hep-2 cells. Conclusion: Proliferation of Hep-2 cells is promoted by MycBCRP which had laid foundation for the further study on BCRP in laryngeal cancer.更多还原