乙肝病毒感染动物模型喜马拉雅旱獭IL-15分子的克隆及序列分析  

Cloning and Sequence Analysis of Interleukin-15 in Marmota himanayana

作　　者：朱彬[1] 朱珍妮[1,2] 李安意[3] 杨雪晟 陶元清[4] 夏幼辰[1] 黄顺梅[1] 宋志韬[1,5] 王忠东[3] 王宝菊[1] 陆蒙吉[6] 杨东亮[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院感染科,武汉430022 [2]华中科技大学同济医学院附属协和医院感染科,武汉430022 [3]湖北省妇幼保健院儿内科,武汉430070 [4]华中科技大学同济医学院附属同济医院实验动物中心,武汉430030 [5]青海省地方病预防控制所实验动物中心,西宁811602 [6]华中科技大学同济医学院附属武汉市中心医院消化科,武汉430014

出　　处：《华中科技大学学报（医学版）》2014年第2期125-129,136,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家传染病科技重大专项课题(No.2008ZX10002011,2012ZX100 04503);国家国际科技合作项目(No.2011DFA31030)

摘　　要：目的对新近建立的乙肝病毒感染动物模型喜马拉雅旱獭的白细胞介素-15(IL-15)分子进行克隆和序列分析。方法抽提喜马拉雅旱獭脾脏总RNA,RT-PCR扩增旱獭IL-15cDNA序列,克隆至pMD18-T载体,制备重组质粒酶切、基因测序鉴定。利用软件分析旱獭IL-15与其他哺乳动物IL-15的同源性、种系亲缘关系,并预测蛋白结构。结果 RT-PCR扩增获得大小为489bp的序列,编码162个氨基酸。旱獭IL-15和土拨鼠IL-15同源性最高,和大鼠同源性最低。种系进化树结果提示旱獭IL-15与土拨鼠IL-15亲缘关系最近。旱獭IL-15蛋白构象与土拨鼠IL-15空间构象非常相似。结论克隆了喜马拉雅旱獭IL-15完整编码序列,为更好地利用新型乙肝病毒感染动物模型喜马拉雅旱獭奠定了基础。Objective To clone and analyze the sequence of interleukin-15（IL-15）in Marmota himanayana,the newly established animal model for the study of hepatitis B virus（HBV）infection.Methods Total RNA was isolated from spleen tissues of Marmota himanayana.cDNA fragments of IL-15in Marmota himanayana was amplified by reverse transcription polymerase chain reaction（RT-PCR）and cloned into the vector pMD18-T.The recombinant plasmids were identified by restricted enzyme digestion and sequenced commercially.The homology of IL-15in Marmota himanayanato other mammal animals,its affinity to other species and the protein structure of IL-15were analyzed by using some software packages.Results Sequence analysis indicated that cDNA fragments（489bp）coding IL-15（162amino acids）was obtained.Homology and phylogenetic analysis indicated that IL-15in Marmota himanayana had the highest homology to that in woodchuck and the lowest homology to that in rat.The homology modeling revealed that the 3Dstructure of IL-15protein in Marmota himanayana was similar to that of woodchuck.Conclusion The complete sequences of IL-15in Marmota himanayana was obtained,which provides a basis for further studying the Chinese Marmot animal model.

关 键 词：喜马拉雅旱獭 白细胞介素-15 结构预测 乙型肝炎病毒 动物模型 

分 类 号：R512.62[医药卫生—内科学]