藏羚羊PGC-1α基因编码区的克隆与分析  

Molecular Cloning and Sequence Analysis of PGC-1α cDNA in Tibetan Antelope

作　　者：马燕[1] 常荣[1] 祁玉娟[1] 格日力[1] 

机构地区：[1]青海大学医学院高原医学研究中心,西宁810001

出　　处：《动物学杂志》2012年第1期25-35,共11页Chinese Journal of Zoology

基　　金：国家973计划项目(No.2012CB518200);国家自然科学基金项目(No.31161861)

摘　　要：以藏羚羊(Pantholops hodgsonii)及同海拔分布的藏系绵羊(Tibetan Sheep)的心肌组织为材料,提取总RNA,利用逆转录聚合酶链反应(RT-PCR)技术扩增出过氧化物酶体增生物激活受体γ辅激活因子-1α(PGC-1α)的基因编码区cDNA片段,与载体连接构建重组质粒,经转化、扩增培养、鉴定后测序。利用生物信息学方法分析显示,藏羚羊和藏系绵羊的PGC-1α基因编码区长度均为2 349 bp,编码797个氨基酸(GenBank登录号分别为:JF449959和JF449960);与其他脊椎动物PGC-1α基因的核苷酸及氨基酸序列相似性达到90%以上;其包含RNA/DNA结合位点、RNA识别基序(RRM)、与核呼吸因子1(NRF-1)及肌细胞增强因子2C(MEF2C)相互作用的区域、富含丝氨酸/精氨酸的结构域、负调节功能结构域、LXXLL模体以及TPPTTPP和DHDYCQ两个保守序列,14个氨基酸差异性位点位于以上部分功能结构域中;此外,磷酸化位点的预测提示藏羚羊可能存在一个潜在的蛋白激酶G的磷酸化位点(第329位的苏氨酸)。本研究成功克隆出了藏羚羊PGC-1α基因的编码区序列,为从能量代谢角度深入探讨藏羚羊适应高原的分子生物学机制提供了新的思路。Total RNAs were extracted from myocardium of Tibetan Antelope（Pantholops hodgsonii） and Tibetan Sheep,both inhabiting on Tibetan Plateau（altitude 4 300 m）.PGC-1α coding cDNA sequences were cloned with reverse transcription polymerase chain reaction（RT-PCR）,and the sequences were confirmed by DNA sequencing.The cloning and sequencing results confirmed that the PGC-1α gene coding sequences of both Tibetan Antelope and Tibetan sheep showed above 90% identity with other species.In addition,the cloned sequences contained the RNA/DNA binding sites,RRM（RNA recognition motif）,the domains involved in the interaction with NRF-1 and MEF2C,Arg/Ser rich domain,negative regulatory domain,LXXLL motif,as well as conserved sequences like TPPTTPP and DHDYCQ,which are present in all PGC-1 family members.Fourteen variable amino acid sites were identified in the functional domains mentioned above.Additionally,analysis of generic phosphorylation sites and kinase specific phosphorylation prediction sites indicated that the 329-threonine amino acid site could be phosphorylated by PKG,which may be unique to Tibetan Antelope.Secondary structures of PGC-1α protein from Tibetan Antelope and Tibetan Sheep were also predicted in this study.In summary,the PGC-1α gene coding regions from Tibetan antelope and Tibetan Sheep have been successfully cloned,which may provide fundamental data for further investigating high altitude adaptation related to genetics in the future.

关 键 词：CDNA克隆 PGC-1Α 能量代谢 藏羚羊 

分 类 号：Q78[生物学—分子生物学]