摘要
目的克隆食管癌组织和癌旁组织中HPV11型主要衣壳蛋白L1基因,并比较两个序列间的同源性。方法以食管癌组织和癌旁组织纯化的总DNA为模板,根据HPV11型L1基因的保守区分段设计引物。分两段扩增HPVL1基因,克隆入质粒载体中,pGEM-3Zf(-)以双脱氧法双向测定目的片段的序列,并拼接出该片段的全序列,比较两个序列间的同源性,以及与已知基因序列的差异性。结果从1例食管癌患者食管癌组织和癌旁组织中,分别克隆到1株HPV11型L1的编码序列M1和M2,两序列间的同源性极高,仅在个别位点存在差异。与GenBank中登录的HPV序列NC001525.1(HPV-11)、M14119.1(HPV-11)和AF217526.1(HPV-11)相比较大部分相同。结论成功地构建了HPV11型L1基因上游片段pGEM-3Zf(-)和下游片段pGEM-3Zf(-)的重组克隆,为深入研究HPV的感染与食管癌发病机制的关系奠定了基础。
Aim To clone major capsid L1 gene of human popillomavirus in esophageal carcinoma and paracancerous tissues and to compare their homology. Methods The DNA purified from the cancerous and paracancerous tissues as template, we designed the primers according to the relatively reserved region in L1 gene. The DNA fragment was cloned into plasmid pGEM 3Zf( ), sequenced by autosequencing instrument from two directions and then compared with those having the corresponding region of HPV. Results A clone of HPV 11 major capsid L1 gene was obtained from two tissues. There was high homology between them except individual site. By comparison, other HPV 11 sequences from GenBank, including NC001525.1(HPV 11), M14119.1(HPV 11) and AF217526.1(HPV 11), also possessed high homology. Conclusion Two recombined clones of HPV 11 major capsid L1 gene are obtained successfully and lay the foundation for the further study on relationship between HPV infection and pathogenesis of esophageal carcinoma.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第3期251-254,共4页
Chinese Journal of Cellular and Molecular Immunology
