论文部分内容阅读
本实验采用ＰＣＲ方法，以中国妇女宫颈癌组织ＤＮＡ为模板扩增出了３株ＨＰＶ１６Ｌ１晚期基因ＤＮＡ片段，扩增参数为９４℃１ｍｉｎ，５５℃３０ｓ，７２℃２ｍｉｎ，共３５个循环。将扩增产物ＨＰＶ１６Ｌ１ＤＮＡ片段与克隆载体ｐＣＲ２．１连接构建了ＨＰＶ１６Ｌ１－ｐＣＲ２．１重组质粒。经酶切ＨＰＶ１６Ｌ１－ｐＣＲ２．１重组质粒回收ＨＰＶ１６Ｌ１基因片段，将其与表达载体ｐＰＩＣ３．５连接，构建了ＨＰＶ１６Ｌ１－ｐＰＩＣ３．５重组体，并经酶切鉴定得到确认。为进一步对感染型ＨＰＶ１６Ｌ１晚期基因的测序及表达打下了基础。
In this study, PCR method was used to amplify three HPV16L1 late-stage DNA fragments from Chinese women’s cervical cancer tissue DNA. The amplification parameters were 94 ℃ for 1min, 55 ℃ for 30s, 72 ℃ for 2min for 35 cycles. The HPV16 L1 DNA fragment was ligated with the cloning vector pCR2.1 to construct the HPV16 L1-pCR2.1 recombinant plasmid. HPV16L1-pCR2.1 recombinant plasmids were recovered HPV16L1 gene fragment, and the expression vector pPIC3.5 ligated to construct HPV16L1-pPIC3.5 recombinant, and identified by restriction endonuclease confirmed. Which lays the foundation for the further sequencing and expression of the late-stage HPV16 L1 gene.