CAJ | 학술논문

目的克隆与肥胖相关的人神经肽Y(NPY)基因,并鉴定该克隆基因序列的正确性.方法根据GeneBank中收录的NPY基因序列,设计该基因上下游序列,经过PCR反应合成NPY的cDNA.然后与pET28a+载体重组,筛选阳性克隆和DNA序列分析鉴定,测序后与GeneBank中NPY基因进行同源性比较和序列分析.结果 PCR扩增出一个100 bp左右的DNA片段,与载体重组后和DNA序列分析鉴定,DNA序列分析显示克隆的DNA片段是人NPY基因.且所克隆的基因共编码36个氨基酸,分子量为4.2 kD,与GeneBank中NPY基因序列同源性达100%.结论克隆的人NPY基因与Genebank中NPY序列完全一致,为进一步应用分子生物学技术深人研究NPY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.
목적 극륭여비반상관적인신경태Y(NPY)기인,병감정해극륭기인서렬적정학성.방법 근거GeneBank중수록적NPY기인서렬,설계해기인상하유서렬,경과PCR반응합성NPY적cDNA.연후여pET28a+재체중조,사선양성극륭화DNA서렬분석감정,측서후여GeneBank중NPY기인진행동원성비교화서렬분석.결과 PCR확증출일개100 bp좌우적DNA편단,여재체중조후화DNA서렬분석감정,DNA서렬분석현시극륭적DNA편단시인NPY기인.차소극륭적기인공편마36개안기산,분자량위4.2 kD,여GeneBank중NPY기인서렬동원성체100%.결론 극륭적인NPY기인여Genebank중NPY서렬완전일치,위진일보응용분자생물학기술심인연구NPY여인체비반발생、발전급전화등상관작용궤제급응용해기인진행기기인단백표체전정료기출.
Objective To clone human neuropeptide Y (NPY),one of the human obesity-related genes,and identifying the sequence of the cloned gene.MethodsUpstream and downstream gene sequence is designed according to NPY gene sequence reported in Genebank.The cDNA of NPY gene is acquired by reaction with PCR.The PCR products are recombined with pET28a + vector,positive clones are selected and DNA sequence analysis is conducted for the identification,followed by sequence analysis and homology comparison with that reported in Genebank.Results A DNA fragment of about 100bp was obtained,the cDNA sequence of the recombinant plasmid of pET28a + with the fragment was analyzed,and the result shows that the cloned DNA fragment is just human NPY cDNA.The cloned gene encodes 36 amino acids with a molecular weight of 42 kD.The nucleotide sequence homology of.the cloned NPY is 100% in comparison with that reported in Genebank.Conclusion The sequence of NPY cDNA that we have successfully cloned is the same as that of NPY in Genebank,which lays the foundation for further studies of gene expression of NPY and the interrelationship of NPY and the genesis,development and transformation of obesity by means of molecular biology.