中间锦鸡儿CiMYB60基因克隆及表达分析

Cloning and Expression Analysis of CiMYB60 Gene from Caragana intermedia

本研究以中间锦鸡儿作为实验材料,利用PCR技术分别对CiMYB60基因的cDNA与gDNA进行了克隆。测序结果表明:CiMYB60基因的开放阅读框为1 035 bp,编码345个氨基酸,其基因组DNA序列长度为1 485 bp,含有2个内含子和3个外显子。生物信息学分析显示:CiMYB60所编码蛋白的N端包含2个MYB结构域,因此认为其属于典型的R2R3-MYB类蛋白。克隆得到CiMYB60基因的ATG上游序列1 848 bp,分析显示该启动子中包含一些与光反应、组织特异性表达、激素和非生物胁迫相关的响应元件。对CiMYB60基因的表达模式进行分析发现在脱水、NaCl和UV-B处理下其表达量随胁迫处理时间的延长而降低。上述研究结果表明CiMYB60基因可能参与中间锦鸡儿对非生物胁迫等逆境胁迫的响应过程,为进一步研究该基因在非生物胁迫中的功能和表达调控提供了一定的参考。

In this study, C. in termedia was used as materials, and the cDNA and gDNA of CiMYB60 gene was cloned by PCR technique. Sequencing results indicate that CiMYB60 had an open reading frame of 1 035 bp,encoding 345 amino acids. The full-length gDNA of CiMYB60 was 1 485 bp, including two introns and three exons. Bioinformatics analysis showed that N-terminal of CiMYB60 contained two MYB domains, so it was regarded as a typical R2 R3-MYB protein. 1 848 bpATG upstream sequence of CiMYB60 was obtained. Analysis showed that the promoter contained some response elements related to light response, tissue specific expression,hormone and abiotic stress. The expression pattern of CiMYB60 was analyzed, which revealed the expression of it was decreased with the prolongation of dehydration, NaCl and UV-B stress. These results demonstrated that CiMYB60 might be involved in the response of of C. intermedia to abiotic stress, which could provide reference for further study on the function and expression regulation of the gene in abiotic stress.