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作 者:胡仁建[1] 吴旭峰[1] 徐涛[1] 沈刘[1] 殷大德[1]
机构地区:[1]重庆理工大学药学与生物工程学院,重庆400054
出 处:《重庆理工大学学报(自然科学)》2011年第10期26-29,共4页Journal of Chongqing University of Technology:Natural Science
基 金:重庆市教委科技项目(KJ110820)
摘 要:培养宫颈癌细胞株HeLa细胞(含HPV18型)并提取其DNA,以用于HPV18E6、E7等基因的构建和表达。复苏HeLa细胞,用含10%血清的DMEM培养基培养、传代、收集离心细胞,用组织/细胞DNA抽提试剂盒提取HeLa细胞的DNA,用核酸电泳进行检验。结果显示:HeLa细胞生长良好,核酸电泳显示有一条与HeLa的DNA大小一致的片段。因宫颈癌HeLa细胞含HPV18型,HeLa细胞的培养及DNA的成功提取为后续的HPV18型E6、E7等基因的构建与表达做准备,从而为宫颈癌的预防、早期诊断和早期治疗奠定基础。For construction and expression of the E6 and E7 genes of HPV18 type,we cultured the HeLa cells (including HPV18 type) of the cervical cancer cell line and extracted the DNA. The HeLa cells were recovered and cultured in DMEM medium containing 10% calf serum,then passed after a few generations ,until we get enough of them. The HeLa cells were collected by centrifugation after that the cells' DNA was extracted by tissue/cell DNA extraction kit, and tested by agarose gel electrophoresis. The results show that the growth of HeLa cells is good,besides,the nucleic acid electrophoresis displays a fragment of the extracting DNA that is consistent with the size of HPV18 type of HeLa cells DNA. Because of cervical cancer HeLa cells containing HPV18 type,the success of HeLa cells culture and DNA extraction can make preparations for the follow-up of E6 and E7 genes construction and expression of type HPV18. Thus it lays a foundation for cervical cancer prevention,early diagnosis and treatment.
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