中国医科大学学报
中國醫科大學學報
중국의과대학학보
JOURNAL OF CHINA MEDICAL UNIVERSITY
2015年
2期
123-126
,共4页
封瑞%刘彦%杨磊%胡慧媛%郭凤%赵美眯%赵金生%郝丽英
封瑞%劉彥%楊磊%鬍慧媛%郭鳳%趙美瞇%趙金生%郝麗英
봉서%류언%양뢰%호혜원%곽봉%조미미%조금생%학려영
钙调蛋白%基因克隆%序列%编码区
鈣調蛋白%基因剋隆%序列%編碼區
개조단백%기인극륭%서렬%편마구
calmodulin%gene cloning%sequence%coding region
目的:克隆豚鼠钙调蛋白2(CaM2)基因编码区及3′端非编码区,为豚鼠CaM基因功能等研究提供遗传学信息。方法以豚鼠心肌组织作为实验材料提取RNA,通过RT?PCR和3′?RACE PCR的方法扩增CaM2的编码区和3′端非编码区序列,应用基因工程技术插入克隆载体构建重组质粒后基因测序及分析。结果克隆出豚鼠CaM2基因编码区450 bp序列和3′端非编码区660 bp序列。分析编码区序列所编码的氨基酸序列与其他种属其他亚型完全一致。而3′端非编码区与其他亚型的同源性较低。结论成功获得了豚鼠CaM2基因编码区和3′端非编码区序列,为进一步研究CaM2的基因功能及其在相关疾病中的作用奠定基础。
目的:剋隆豚鼠鈣調蛋白2(CaM2)基因編碼區及3′耑非編碼區,為豚鼠CaM基因功能等研究提供遺傳學信息。方法以豚鼠心肌組織作為實驗材料提取RNA,通過RT?PCR和3′?RACE PCR的方法擴增CaM2的編碼區和3′耑非編碼區序列,應用基因工程技術插入剋隆載體構建重組質粒後基因測序及分析。結果剋隆齣豚鼠CaM2基因編碼區450 bp序列和3′耑非編碼區660 bp序列。分析編碼區序列所編碼的氨基痠序列與其他種屬其他亞型完全一緻。而3′耑非編碼區與其他亞型的同源性較低。結論成功穫得瞭豚鼠CaM2基因編碼區和3′耑非編碼區序列,為進一步研究CaM2的基因功能及其在相關疾病中的作用奠定基礎。
목적:극륭돈서개조단백2(CaM2)기인편마구급3′단비편마구,위돈서CaM기인공능등연구제공유전학신식。방법이돈서심기조직작위실험재료제취RNA,통과RT?PCR화3′?RACE PCR적방법확증CaM2적편마구화3′단비편마구서렬,응용기인공정기술삽입극륭재체구건중조질립후기인측서급분석。결과극륭출돈서CaM2기인편마구450 bp서렬화3′단비편마구660 bp서렬。분석편마구서렬소편마적안기산서렬여기타충속기타아형완전일치。이3′단비편마구여기타아형적동원성교저。결론성공획득료돈서CaM2기인편마구화3′단비편마구서렬,위진일보연구CaM2적기인공능급기재상관질병중적작용전정기출。
Objective To clone the coding region and 3′non?coding region of calmodulin 2(CaM2)in guinea pig,to provide the genetic informa?tion for studying the gene function of Calmodulin 2. Methods Total RNA was extracted from heart tissue of guinea pig,the coding region and 3′non?coding region of CaM2 were amplified by RT?PCR and 3′?RACE PCR methods,and the recombinant plasmid was constructed by inserting cDNA of the coding region and 3′non?coding region of CaM2 into the cloning vector by genetic engineering technology followed by DNA sequencing and se?quence analysis. Results The cloned coding region of CaM2 was 450 bp,and the 3′non?coding region of CaM2 was 660 bp. The amino acid se?quences of the coding region of CaM2 was consistent with those of other CaM subtypes,and the 3′non?coding region of CaM2 had low homology with those of other subtypes. Conclusion The cloning of CaM2 coding region and 3′non?coding region in guinea pig was the foundation for further study on the gene function of CaM2 and its role in related diseases.