摘要
目的克隆豚鼠钙调蛋白2(Ca M2)基因编码区及3′端非编码区,为豚鼠Ca M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Ca M2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Ca M2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Ca M2基因编码区和3′端非编码区序列,为进一步研究Ca M2的基因功能及其在相关疾病中的作用奠定基础。
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 eDNA 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.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2015年第2期123-126,共4页
Journal of China Medical University
基金
国家自然科学基金(31071004
31471091
31400981)
关键词
钙调蛋白
基因克隆
序列
编码区
calmodulin
gene cloning
sequence
coding region
