国际护理学杂志
國際護理學雜誌
국제호이학잡지
INTERNATIONAL JOURNAL OF NURSING
2009年
4期
557-560
,共4页
丁克祥%董萍%郑永晨%杨永鹏%朱晓亮%韩晋云%单志新%丁宇%丁振华
丁剋祥%董萍%鄭永晨%楊永鵬%硃曉亮%韓晉雲%單誌新%丁宇%丁振華
정극상%동평%정영신%양영붕%주효량%한진운%단지신%정우%정진화
肥胖%神经肽Y受体Y1%基因克隆%DNA序列分析
肥胖%神經肽Y受體Y1%基因剋隆%DNA序列分析
비반%신경태Y수체Y1%기인극륭%DNA서렬분석
Obesity%Neuropeptide Y(NPY)receptor Y1%Gene cloning%DNA sequence analysis
目的 克隆与肥胖相关的人神经肽Y受体Y1(NPYlR)基因,并鉴定该克隆基因序列的正确性.方法 从人的脂肪组织中提取总RNA并进行反转录,以此为模板用PCR扩增NPY1R基因的cDNA,然后与pET 28a+载体重组,筛选阳性克隆和DNA序列分析鉴定,测序后与GeneBank中NPY1R基因进行同源性比较和序列分析.结果 PER扩增出一个1100 bp左右的DNA片段,与载体重组后DNA序列分析鉴定,DNA序列分析显示克隆的DNA片段是人NPY1R基因,且所克隆的基因共编码384个氨基酸,分子量为44 KD,与GeneBank中NPY1R基因序列同源性达100%.结论 克隆的人NPY1R基因与GeneBank中NPY1R序列完全一致,为进一步应用分子生物学技术深入研究NPY1R与人体肥胖发生、发展及转化等相关作用机制及应用该基因进行其基因蛋白表达奠定了基础.
目的 剋隆與肥胖相關的人神經肽Y受體Y1(NPYlR)基因,併鑒定該剋隆基因序列的正確性.方法 從人的脂肪組織中提取總RNA併進行反轉錄,以此為模闆用PCR擴增NPY1R基因的cDNA,然後與pET 28a+載體重組,篩選暘性剋隆和DNA序列分析鑒定,測序後與GeneBank中NPY1R基因進行同源性比較和序列分析.結果 PER擴增齣一箇1100 bp左右的DNA片段,與載體重組後DNA序列分析鑒定,DNA序列分析顯示剋隆的DNA片段是人NPY1R基因,且所剋隆的基因共編碼384箇氨基痠,分子量為44 KD,與GeneBank中NPY1R基因序列同源性達100%.結論 剋隆的人NPY1R基因與GeneBank中NPY1R序列完全一緻,為進一步應用分子生物學技術深入研究NPY1R與人體肥胖髮生、髮展及轉化等相關作用機製及應用該基因進行其基因蛋白錶達奠定瞭基礎.
목적 극륭여비반상관적인신경태Y수체Y1(NPYlR)기인,병감정해극륭기인서렬적정학성.방법 종인적지방조직중제취총RNA병진행반전록,이차위모판용PCR확증NPY1R기인적cDNA,연후여pET 28a+재체중조,사선양성극륭화DNA서렬분석감정,측서후여GeneBank중NPY1R기인진행동원성비교화서렬분석.결과 PER확증출일개1100 bp좌우적DNA편단,여재체중조후DNA서렬분석감정,DNA서렬분석현시극륭적DNA편단시인NPY1R기인,차소극륭적기인공편마384개안기산,분자량위44 KD,여GeneBank중NPY1R기인서렬동원성체100%.결론 극륭적인NPY1R기인여GeneBank중NPY1R서렬완전일치,위진일보응용분자생물학기술심입연구NPY1R여인체비반발생、발전급전화등상관작용궤제급응용해기인진행기기인단백표체전정료기출.
Objective To clone the human nettropeptide Y receptor Y1(NPY1R),which is one of the human obesity-related genes,and identify the cloned gene sequence.Methods The cDNA of NPY1R gene was amplified from RNA of the human fat by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were recombined with pET28a+vector,the positive clones were selected and DNA sequence analysis was conducted for the identification.followed by sequence analysis and homology comparison with that reported in Genebank after sequencing.Results A DNA fragment of about 1100bp was obtained by RT-PCR,the cDNA sequence of the recombinant plasmid of pET28a+with the fragment was analyzed,and the result showed that the cloned DNA fragment was just human NPY1R cDNA.The cloned gene encoded 384 amino acids with a molecular weight of 44KD.The nucleotide sequence homology of the cloned NPY1R was 100% in comparison with that reported in Genebank.Conclusions The sequence of NPY1R cDNA cloned successfully is the same as that of NPY1R in Genehank,which lays the foundation for further studics of gene expression of NPY1R and the interrelationship between NPY1R and genesis,development and transformation of obesity by nlesns of molecular biology.
