中间锦鸡儿CiCML23基因克隆及序列分析

Cloning and Sequence Analysis of CiCML23 Gene from Caragana intermedia

本研究以抗旱、抗寒、耐盐碱的中间锦鸡儿为材料,筛选其优良抗逆基因,为林草基因工程提供基因源。在中间锦鸡儿干旱胁迫转录组文库中找到1条注释为CiCML基因的序列,设计引物对其进行克隆。克隆得到CiCML基因cDNA序长列817 bp,包括426 bp开放阅读框,编码142个氨基酸。通过比对分析发现该基因与豆科模式植物蒺藜苜蓿CML23基因同源,具有较近的亲缘关系,因此命名为CiCML23。预测其等电点为4.42,分子质量为15.65 kD,是稳定的亲水性蛋白,存在CML典型的EF-hand结构域。初步阐明CiCML23基因的序列特征,为进一步预测CiCML23基因的功能提供参考依据。

In this study, the Caragana intermedia, which is resistant to drought, cold and salt, was used as a material to screen its excellent stress-resistant genes, providing a genetic source for genetic engineering of forest and forage species. A sequence annotated with the CiCML gene was found from the transcriptome library constructed in our laboratory using drought stressed seedlings of the Caragana intermedia and primers were designed to clone it. The cDNA sequence of the CiCML gene was 817 bp long, including a 426 bp open reading frame which encoding 142 amino acids. The gene was found to be homologous to the leguminous plant Medicago truncatula CML23 gene by comparison analysis, and was named as CiCML23. The isoelectric point is predicted to be pI of 4.42 and the molecular mass is 15.65 kD. It is a stable hydrophilic protein with the typical EF-hand domain of CML. The sequence characteristics of CiCML23 gene were preliminarily elucidated, which provide reference basis for further prediction of the function of CiCML23 gene.