豚鼠钙调蛋白2基因编码区及3′端非编码区序列克隆

摘要

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.%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。