华中科技大学学报（医学版）
華中科技大學學報（醫學版）
화중과기대학학보（의학판）
ACTA UNIVERSITATIS MEDICINAE TONGJI
2014年
2期
125-129,136
,共6页
朱彬%朱珍妮%李安意%杨雪晟%陶元清%夏幼辰%黄顺梅%宋志韬%王忠东%王宝菊%陆蒙吉%杨东亮
硃彬%硃珍妮%李安意%楊雪晟%陶元清%夏幼辰%黃順梅%宋誌韜%王忠東%王寶菊%陸矇吉%楊東亮
주빈%주진니%리안의%양설성%도원청%하유신%황순매%송지도%왕충동%왕보국%륙몽길%양동량
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
喜馬拉雅旱獺%白細胞介素-15%結構預測%乙型肝炎病毒%動物模型
희마랍아한달%백세포개소-15%결구예측%을형간염병독%동물모형
Marmota himanayana%interleukin-15%structure prediction%hepatitis B virus%animal model
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-15 cDNA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489 bp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。
目的：對新近建立的乙肝病毒感染動物模型喜馬拉雅旱獺的白細胞介素-15（IL-15）分子進行剋隆和序列分析。方法抽提喜馬拉雅旱獺脾髒總 RNA ，RT-PCR擴增旱獺IL-15 cDNA序列，剋隆至pMD18-T載體，製備重組質粒酶切、基因測序鑒定。利用軟件分析旱獺IL-15與其他哺乳動物IL-15的同源性、種繫親緣關繫，併預測蛋白結構。結果 RT-PCR擴增穫得大小為489 bp的序列，編碼162箇氨基痠。旱獺IL-15和土撥鼠IL-15同源性最高，和大鼠同源性最低。種繫進化樹結果提示旱獺IL-15與土撥鼠IL-15親緣關繫最近。旱獺IL-15蛋白構象與土撥鼠IL-15空間構象非常相似。結論剋隆瞭喜馬拉雅旱獺IL-15完整編碼序列，為更好地利用新型乙肝病毒感染動物模型喜馬拉雅旱獺奠定瞭基礎。
목적：대신근건립적을간병독감염동물모형희마랍아한달적백세포개소-15（IL-15）분자진행극륭화서렬분석。방법추제희마랍아한달비장총 RNA ，RT-PCR확증한달IL-15 cDNA서렬，극륭지pMD18-T재체，제비중조질립매절、기인측서감정。이용연건분석한달IL-15여기타포유동물IL-15적동원성、충계친연관계，병예측단백결구。결과 RT-PCR확증획득대소위489 bp적서렬，편마162개안기산。한달IL-15화토발서IL-15동원성최고，화대서동원성최저。충계진화수결과제시한달IL-15여토발서IL-15친연관계최근。한달IL-15단백구상여토발서IL-15공간구상비상상사。결론극륭료희마랍아한달IL-15완정편마서렬，위경호지이용신형을간병독감염동물모형희마랍아한달전정료기출。
Objective To clone and analyze the sequence of interleukin-15 (IL-15) in Marmota himanayana ,the newly es-tablished 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-15 in 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-15 in Marmota himanayana to other mammal animals ,its affinity to other species and the protein structure of IL-15 were analyzed by using some software packages.Results Sequence analysis in-dicated that cDNA fragments (489 bp) coding IL-15 (162 amino acids) was obtained.Homology and phylogenetic analysis indi-cated that IL-15 in Marmota himanayana had the highest homology to that in woodchuck and the lowest homology to that in rat.The homology modeling revealed that the 3D structure of IL-15 protein in Marmota himanayana was similar to that of woodchuck.Conclusion The complete sequences of IL-15 in Marmota himanayana was obtained ,which provides a basis for further studying the Chinese Marmot animal model.
