拟南芥钴胺素合成相关蛋白CSRP基因克隆及其功能分析

以拟南芥为研究对象,根据已知的生物信息数据,通过反向遗传学的方法,对与钴胺素合成相关蛋白CSRP基因进行分子克隆,得到该基因的基因组序列和cDNA序列,构建二元重组质粒,利用根癌农杆菌介导的花浸泡转基因方法分别转化突变体和野生型植株,进行互补试验和过表达试验,筛选转基因植株,并进一步预测其编码蛋白的亚细胞定位于叶绿体中.结果表明,在突变体中转入该基因后,转基因植株恢复野生型的表型,而在过表达的转基因植株中表型更为明显,即生长旺盛,晚花.说明该基因对于拟南芥的营养生长起促进作用,由于其功能预测与钴胺素合成相关的蛋白,推测其可能在植物营养生长中起作用.

Cloning of a cobalamin synthesis related gene and its predicted function analysis in Arabidopsis thaliana

A gene which encodes the cobalamin synthesis related protein in Arabidopsis thaliana was sdudied with the reserve genetic methods according to the bioinformation knowledge,and the predicted protein function was been further analysis.We have cloned the gene and obtained the genomic DNA and cDNA.The binary recombinant were constructed and transformed into knockout mutant or wide type Arabidopsis respectively by Agrobacterum-mediated transformation through floral-dipping method for complementation and overexpression experiment.The transgenic plants were screened.Further more,the subcellular location and the function of the encoded protein was predicted.The results show that the gene can rescue the mutant phenotype to wide type,and the overexpressed transgenic plants become strong and late followed.So the gene has something function with the vegetative growth,because of the predicting structure and function has something do with the cobalamin synthesis,it is suggested that the cobalamin may play a role in the vegetative growth of the plants.