蒺藜苜蓿B561基因的克隆及对拟南芥的转化

Cloning of B561 Gene from Medicago truncatula and Transformation of Arabidopsis thaliana

细胞色素B561是一种负责电子传递的整膜蛋白,在植物生长发育、抗毒素防御反应方面具有重要作用,并且能够防止植物在干旱条件下由于过度光照而受损。本研究以蒺藜苜蓿为试验材料,采用RACE技术获得蒺藜苜蓿B561基因全长,其编码区长807 bp,编码269个氨基酸。生物信息学分析显示:该基因编码蛋白与其他物种B561蛋白具有较高的同源性,与大豆的同源性最高。利用DNA重组技术将B561基因完整编码区连接至基因表达载体pBI121,成功构建植物表达载体pBI-B561,并通过农杆菌介导法转将其转化至拟南芥,成功获得25株转基因植株。本研究为进一步探究B561基因的功能提供了数据参考和试验材料。

Cytochrome B561 is a whole membrane protein which is responsible for electron transport.It plays an important role in plant growth and development,as well as anti-toxin defense response,and can prevent plants from being damaged by excessive light under drought conditions.In this study,the Medicago truncatula B561 gene was obtained by RACE technology.The coding domain was 807 bp,encoding 269 amino acids.Bioinformatics analysis showed that protein coding by this gene had high homology with other species B561 protein and the highest homology with Glycine max L..The complete coding region of B561 gene was connected to plant expression vector pBI121 by DNA recombination technology,and the plant expression vector pbi-B561 was successfully constructed,the plant over-expression vector of pBI-B561 was successfully constructed,transferring into Arabidopsis by Agrobacterium-mediated method.PCR results showed that B561 gene had been successfully integrated into the genome of Arabidopsis thaliana,and 25 transgenic plants were successfully obtained.This study provides data reference and experimental materials for further exploring the function of B561 gene.