中间锦鸡儿CiMYB68基因克隆及表达分析  

Cloning and Expression Analysis of CiMYB68 from Caragana intermedia

作　　者：冯宗琪 韩晓敏[1] 杨杞[1] 邢丹丹[1] 齐力旺[2] 王瑞刚[1] 李国婧[1] 

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018 [2]中国林业科学院林业研究所,北京100091

出　　处：《西北植物学报》2014年第6期1083-1089,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31360056);教育部高校博士学科点基金(20121515110007);国家高技术研究发展计划(2011AA100203)

摘　　要：该研究以中间锦鸡儿(Caragana intermedia)为材料,利用RACE技术克隆了CiMYB68基因的全长序列。对CiMYB68的基因组DNA和cDNA全长分析显示,CiMYB68基因无内含子,开放阅读框为852bp,编码284个氨基酸。预测CiMYB68基因编码的蛋白质等电点为8.95,分子量约为31 459.4Da。序列比对和系统进化分析表明,该蛋白和大豆的GmMYB68一致性最高,达到67%。构建了CiMYB68基因与GFP融合表达质粒,激光共聚焦显微镜观察发现,融合蛋白定位于细胞核。用实时荧光定量PCR技术对在不同胁迫条件下CiMYB68基因的表达检测结果表明,在干旱和低温处理下CiMYB68均受到不同程度的诱导,暗示CiMYB68基因可能与中间锦鸡儿响应逆境胁迫有关。A MYB encoding gene was cloned by RACE（rapid amplification of cDNA ends） technique from Caragana intermedia.The gDNA and full-length cDNA sequence analysis revealed that this gene contains no intron.The full length ORF was 852 bp,and the deduced protein comprised 284 amino acids with a calculated molecular weight of 31 459.4 Da,as well as an isoelectric point of 8.95.Sequence alignment showed that this MYB protein is relatively close to GmMYB68,with an identity of 67%,so the gene was named as CiMYB68.CiMYB68 and GFP fusion vector was constructed,and GFP fluorescence was observed in the nuclei under confocal laser scanning microscope.Real-time quantitative PCR analysis showed that the transcript of CiMYB68 was induced strongly under drought and cold treatment.These results indicated that CiMYB68 might be involved in stress responses of C.intermedia.

关 键 词：中间锦鸡儿 MYB 基因克隆 表达分析 

分 类 号：Q785[生物学—分子生物学] Q786