论文部分内容阅读
宫颈癌与人乳头瘤病毒感染密切相关,建立宫颈癌实验模型可为宫颈癌的研究提供理想的模拟实验条件。我们通过应用基因重组技术,分别以HPV31型E6和E7基因为目的基因,通过原核表达、蛋白纯化和免疫小鼠等获得其特异性检测抗体。我们还通过构建E6和E7基因真核表达载体、转染C33A细胞、博莱霉素抗性筛选和表达检测等步骤,获得一种稳定的体外宫颈癌细胞系。经酶切鉴定及测序证实细胞基因组已重组插入质粒中的目的基因。我们已成功筛选到稳定的目的mRNA和蛋白表达的阳性细胞系,建立了稳定的人乳头状瘤病毒31型(HPV31)的宫颈癌细胞株,为研究宫颈癌提供了体外实验模型。
Cervical cancer is closely related to human papillomavirus (HPV) infection. Establishing a cervical cancer experimental model can provide ideal simulated experimental conditions for the study of cervical cancer. By using gene recombination technology, we used HPV31 E6 and E7 genes as the target genes respectively and obtained their specific detection antibodies through prokaryotic expression, protein purification and immunization of mice. We also obtained a stable in vitro cervical cancer cell line by constructing E6 and E7 eukaryotic expression vector, transfection of C33A cells, selection and expression detection of bleomycin resistance and other steps. Identified and sequenced by restriction enzyme confirmed that the genome of the cell has been recombined into the target gene in the plasmid. We have successfully screened stable expression of the target mRNA and protein expression of the positive cell lines, the establishment of a stable human papillomavirus type 31 (HPV31) cervical cancer cell lines for the study of cervical cancer provides an experimental model in vitro.