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自我剪切锤头状核酶体外切割HPV11/E2的研究

Studies on Cleavage of HPV11/E2 Gene by Self-cleaved Hammerhead Ribozyme

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【作者】 侯化; 杨光彩; 黄杨中; 王曙;

【Author】 HOU Hua 1,YANG Guang cai 1,HUANG Yang zhong 2,WANG Shu 3 (1.The First Military Medical University,Guangzhou 510515,China;2.University of Virginia,USA 96102; 3.Institute of Genetics,School of Life Science,Fudan University,Shanghai

【机构】 广州同和第一军医大学生化教研室; 美国弗吉尼亚大学生物部; 上海复旦大学生命科学院遗传学研究所 广州501515; 广州501515; 美国96102; 上海200092;

【Abstract】 In order to study the pathogenesis of human papilloma virus type 11 (HPV11) and seek for a therapeutic approach of the disease caused by HPV11,we amplified the HPV11/E2 of 644bp length by PCR with HPV11 plasmid DNA,pGEM T Easy was used as vector and a clone pTV 644 was obtained.The sequencing of inserted DNA was carried out after selecting and identification,according to the hammerhead structure described by Symon’s. We used computer to analyze the possible secondary cleavage sites on HPV11/E2 mRNA and to predict the secondary structure of substrate and ribozyme to exclude the analogous sequence of substrate combined with ribozyme found in the mRNA.The hammerhead ribozyme of RZ3281 against HPV11/E2 mRNA were selected to carry out cleavage reaction in vitro. Results of the experiment showed that 644bp substrate derived from HPV11/E2 can be cleaved site specifically by ribozyme in vitro,cleavage activity showed over 85% under the optimum reaction condition.Self cleavage hammerhead ribozyme can remove effects of gene joint transcript from the vector, that promise the consistent efficiency of ribozyme in vitro and in vivo as far as possible.Results of the experiment demonstrated that the ribozyme will become a highly effective and specific therapy against HPV11 infection.

【关键词】 人乳头瘤病毒11型; E2基因; 核酶; 体外剪切;
【Key words】 HPV11; E2 gene; ribozyme; cleavage reaction in vitro;
【基金】 国家自然科学基金 (3 970 0 187);广东省自然科学基金(970 3 3 2 )
  • 【文献出处】 病毒学报 ,Chinese Journal of Virology , 编辑部邮箱 ,2001年03期
  • 【分类号】Q753
  • 【下载频次】78
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