鸡贫血病毒Apoptin蛋白编码基因的克隆和序列分析  

Cloning and sequencing of Apoptin gene in chicken anaemia virus

作　　者：刘德纯[1] 王健生[1] 王作仁[1] 段小艺[1] 陈武科[1] 杨广笑 

机构地区：[1]西安交通大学医学院第一附属医院肿瘤外科,陕西西安710061 [2]西安华广生物工程公司,陕西西安710025

出　　处：《第四军医大学学报》2007年第11期961-961,962,963,共3页Journal of the Fourth Military Medical University

基　　金：高等学校博士学科点专项科研基金(20060698055)

摘　　要：目的:为了寻找肿瘤特异凋亡蛋白,克隆鸡法氏囊组织中鸡贫血病毒Apoptin蛋白的编码基因并进行序列分析.方法:根据基因数据库资料设计合成引物,以成鸡法氏囊组织中提取的DNA作模板,通过PCR获得了特异扩增产物,将其插入到pGEMT-easy载体并转染感受态E.coli-DH5α菌株,提取质粒经限制性内切酶酶切鉴定后,使用DNA测序仪测定插入片段的DNA序列.使用DNAsis分析软件分析所克隆的DNA序列及其编码蛋白质,并与数据库中Apoptin蛋白的编码基因相比较.结果:序列分析表明,成功地克隆了鸡贫血病毒的Apoptin基因,同基因数据库中的相关资料比较发现与山东报道的Apoptin基因(GenBank AY171617)的核酸序列一致,同鸡贫血病毒全基因序列中的Apoptin序列(GenBank AF313470)有6个核酸差异.结论:Apoptin编码基因的克隆为特异诱导肿瘤细胞凋亡治疗肿瘤奠定了基础.AIM： To clone the chicken anemia virus-derived Apoptin gene from the chicken＇s bursa of Fabricius and to do a sequencing analysis on it. METHODS ： A pair of primers was constructed and synthesized according to the sequences in the Gen- Bank database. The DNA template was isolated from the chicken＇s bursa of Fabficius, amplified specifically by PCR and inserted into the vector of pGEMT-easy. The recombinant vector was used to infect E. coli DH5α cells. The isolated vector was confirmed with restriction endonuclease digestion and the sequence was assessed by DNA sequencer. The DNAsis software was employed to detect the consistence between our product and the one from GenBank. RESULTS： Sequencing analysis indicated that we succeeded in cloning the Apeptin gene. Our sequence was consistent with the reported one in Shandong province （GenBank AY171617）, and different from the GenBank AF313470 in 6 sites. CONCLUSION： Cloning of Apeptin gene may make a foundation for the tumor therapy by inducing specifically tumor cell apoptesis.

关 键 词：鸡贫血病毒 凋亡素 克隆 测序 

分 类 号：R394[医药卫生—医学遗传学]