广东地区HPV16型L1基因的序列分析及其毕赤酵母表达载体的构建  

Sequence analysis of human papillomavirus 16 L1 gene from Guangdong area and construction of its pichia pastories expression vector

作　　者：刘红[1] 宇丽[1] 周羽竝[1] 莫宏波[1] 刘萍 李发涛[1] 李冬艳[1] 罗京资[1] 

机构地区：[1]暨南大学医学院生物化学教研室,广东广州510632 [2]番禺妇幼保健院,广东广州511400

出　　处：《暨南大学学报（自然科学与医学版）》2008年第2期115-120,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：教育部科学技术研究重点项目资助(重点02190);广东省卫生厅医学科研基金资助项目资助(A2001305)

摘　　要：目的:分析广东地区人乳头瘤病毒(HPV)16型L1基因结构特点;构建广东分离株HPV16 L1毕赤酵母分泌型表达载体。方法:采用PCR技术从广东地区宫颈癌组织中扩增HPV16 L1基因,克隆入毕赤酵母表达载体pPICZαC,测序并对HPV16 L1基因进行序列分析。结果:成功扩增了广东地区HPV16 L1基因,并构建了其毕赤酵母表达载体pPICZαC-HPV16 L1。广东分离株HPV16 L1基因序列与德国标准株相比有16处不同,同源性为98.99%,其编码的氨基酸序列有8处发生改变;与中国标准株比较有9处不同,同源性为99.18%,其编码的氨基酸序列有4处发生变化;发现在HPV16 L1基因序列中nt5 649、nt5 652、nt5 654、nt5 657、nt5 692、nt5 693,6个位点存在突变。结论:广东地区HPV16 L1基因序列与德国标准株、中国标准株相比较均存在差异;成功构建广东地区HPV16 L1真核表达载体。Aim：To investigate the structure specificity of human papilloma virus （HPV） type 16 in Guangdong area, and to construct its pichia pastories secretion type expression vector. Methods： The HPV16 L1 gene were amplified from human cervical carcinoma tissues in Guangdong province by PCR, and cloned into pichia pastories expression vector. The HPV16 L1 gene were sequenced and analyzed. Results： The HPV16 L1 gene sequence were obtained, and the HPV16 L1 pichia pastories expression vector were constructed. There were 16 nucleotide differences between Guangdong strain and Germanic standard strain, the sequences were 98.99% homology, subsequently changed 8 amino acids it coded. There were 9 nucleotide differences between Guangdong strain and Chinese standard strain, the sequences were 99. 18% homology, subsequently changed 4 amino acids it coded. There are 6 mutations（nt5 649, nt5 652, nt5 654, nt5 657, nt5 692, nt5 693）in HPV16 L1 gene sequence obtained from Guangdong area. Conclusion： There are nucleotide differences of HPV16 L1 gene among Germanic standard strain, Chinese standard strain and Guangdong strain. Guangdong HPV16 L1 pichia pastories expression vector were constructed.

关 键 词：人乳头瘤病毒16型 广东 L1基因 突变 毕赤酵母 

分 类 号：R373[医药卫生—病原生物学]