四川大学学报(自然科学版)
四川大學學報(自然科學版)
사천대학학보(자연과학판)
JOURNAL OF SICHUAN UNIVERSITY(NATURAL SCIENCE EDITION)
2013年
4期
899-907
,共9页
何夏萍%周彦妮%阎振鑫%李娟%王亚军
何夏萍%週彥妮%閻振鑫%李娟%王亞軍
하하평%주언니%염진흠%리연%왕아군
猪%黑皮质素(Melanocortin)%黑皮质素受体(Melanocortin Receptors)%基因克隆
豬%黑皮質素(Melanocortin)%黑皮質素受體(Melanocortin Receptors)%基因剋隆
저%흑피질소(Melanocortin)%흑피질소수체(Melanocortin Receptors)%기인극륭
pig%melanocortin%melanocortin receptor 3%melanocortin receptor 4%cloning
以长白猪(L andrace)大脑 cDNA 为模板,克隆长白猪黑皮质素受体3和4基因(MC3R和MC4R).序列分析显示:长白猪MC3R基因cDNA 编码具319个氨基酸的前体蛋白,与人、大鼠、小鼠和斑马鱼的氨基酸序列一致性分别为86.4%、83.3%、82%和69.7%;M C4R基因cDN A全长1175 bp ,编码具332个氨基酸的前体蛋白,与人、大鼠、小鼠的氨基酸序列一致性高达94%以上,与斑马鱼M C4 R氨基酸序列享有70.7%的同源性.序列分析显示,猪M C3R和M C4R基因都具有典型的七次跨膜结构域,和短的胞内环和胞外环,且在第五个跨膜域中保守的甲硫氨酸(M )分别替换为异亮氨酸(I )和缬氨酸(V ).另外,在猪M C3 R和M C4R中还鉴定到保守的半胱氨酸残基,第一个胞内环中保守的PM Y基序和N末端的3个N-链接糖基化位点.组织表达图谱分析表明,M C3R主要在大脑和脾中高表达,而M C4R主要在六种检测的组织中都有表达,在肾中表达较弱.
以長白豬(L andrace)大腦 cDNA 為模闆,剋隆長白豬黑皮質素受體3和4基因(MC3R和MC4R).序列分析顯示:長白豬MC3R基因cDNA 編碼具319箇氨基痠的前體蛋白,與人、大鼠、小鼠和斑馬魚的氨基痠序列一緻性分彆為86.4%、83.3%、82%和69.7%;M C4R基因cDN A全長1175 bp ,編碼具332箇氨基痠的前體蛋白,與人、大鼠、小鼠的氨基痠序列一緻性高達94%以上,與斑馬魚M C4 R氨基痠序列享有70.7%的同源性.序列分析顯示,豬M C3R和M C4R基因都具有典型的七次跨膜結構域,和短的胞內環和胞外環,且在第五箇跨膜域中保守的甲硫氨痠(M )分彆替換為異亮氨痠(I )和纈氨痠(V ).另外,在豬M C3 R和M C4R中還鑒定到保守的半胱氨痠殘基,第一箇胞內環中保守的PM Y基序和N末耑的3箇N-鏈接糖基化位點.組織錶達圖譜分析錶明,M C3R主要在大腦和脾中高錶達,而M C4R主要在六種檢測的組織中都有錶達,在腎中錶達較弱.
이장백저(L andrace)대뇌 cDNA 위모판,극륭장백저흑피질소수체3화4기인(MC3R화MC4R).서렬분석현시:장백저MC3R기인cDNA 편마구319개안기산적전체단백,여인、대서、소서화반마어적안기산서렬일치성분별위86.4%、83.3%、82%화69.7%;M C4R기인cDN A전장1175 bp ,편마구332개안기산적전체단백,여인、대서、소서적안기산서렬일치성고체94%이상,여반마어M C4 R안기산서렬향유70.7%적동원성.서렬분석현시,저M C3R화M C4R기인도구유전형적칠차과막결구역,화단적포내배화포외배,차재제오개과막역중보수적갑류안산(M )분별체환위이량안산(I )화힐안산(V ).령외,재저M C3 R화M C4R중환감정도보수적반광안산잔기,제일개포내배중보수적PM Y기서화N말단적3개N-련접당기화위점.조직표체도보분석표명,M C3R주요재대뇌화비중고표체,이M C4R주요재륙충검측적조직중도유표체,재신중표체교약.
In this study ,using cDNA of L andrace pig brain as the template ,the full-length cDNAs of MC3R and MC4R genes have been cloned .Pig MC3R cDNA encodes a protein precursor of 319 amino acids ,which shares high amino acid sequence identity with that of human (86 .4% ) ,rat (83 .3% ) , mouse (82% ) and zebrafish(69 .7% ) .Pig MC4R cDNA is 1175 bp in length and encodes a protein pre-cursor of 332 amino acids ,w hich shares high amino acid sequence identity (>94% ) with that of mam-mals and zebrafish(69 .7% ) .Further structural analyses indicate that pig MC3R and MC4R contain typi-cal seven transmembrane domains and short extracellular (ECL) and intracellular (ICL) loops .The sub-stitution of a methionine for isoleucine (MC3R)/ Valine (MC4R) in the fifth transmembrane domain (TMD 5) also has been identified .In addition ,pig MC3R and MC4R share the PMY motif in the first ICL and 3 conserved N-linked glycosylation sites within N-terminal domain which are conserved in most MCRs .RT-PCR assay revealed that :MC3R is mainly expressed in brain and spleen with high abun-dance .While MC4R is expressed all six pig tissues examined with relatively weak signals in kidney .