关键词: 乳头状瘤病毒，人;病毒壳体;基因扩增;克隆;序列分析

    摘 要:目的 从临床尖锐湿疣标本克隆人乳头瘤病毒(HPV)11型主要衣壳蛋白L1基因，进行序列测定及序列分析比较，以为研究该临床病毒株的免疫原性及流行病学奠定基础. 方法 以临床尖锐湿疣标本总DNA为模板，用L1基因保守区简并引物以PCR方法分段扩增衣壳蛋白L1基因大部，重组入pGEM-3zf(-)质粒载体，以双脱氧法双向测定插入片段序列，拼接出L1基因序列，通过BLAST2.0与已报道序列进行比较. 结果 从西安地区一尖锐湿疣临床标本克隆到的一株HPV11型L1蛋白编码序列与一文献报道大部分相同，但有些位点有点突变. 结论 构建的pGEM-3zf(-)重组质粒为进一步通过杂交、体内分段表达等手段研究该株病毒L1蛋白的免疫学和流行病学性质创造了条件.

    Cloning and sequence analysis of L1major capsid protein gene of human papillomavirus type11

    FENG Yong-Qiang YAN Xiao-Jun SU Cheng-Zhi ZHANG Ju JIANG Hong

    Institute of Genetic Diagnosis of Chinese PLA，Fourth Military Medical University，Xi'an710033，China

    Keywords:papillomavirus，human;capsid;gene amplifica-tion;cloning;sequence analysis

    Abstract:AIM To obtain the L1major capsid protein gene of human papillomavirus type11for the exploring of the im-munological activity and epistemology of L1.METHODS Two over-lapping parts of L1gene were amplified by PCR from the whole DNA extract of clinical specimen of condylo-ma acuminata with the degenerate primers designed within the conserved regions of L1gene.After being cloned into plasmid pGEM-3zf(-)the nucleotide sequence of the inserted amplified fragments was determined by an automated DNA sequencer and then put together to get the whole sequence of L1gene which was analyzed with BLAST2.0against all re-ported sequence afterwards.RESULTS Although the se-quence of the cloned L1gene from this clinical condyloma acuminata specimen in Xi'an was similar to a reported L1gene with an identity of99% ......