拟南芥心皮发育CRABS CLAW基因的克隆与荠菜遗传转化

Cloning of Arabidopsis thaliana CRABS CLAW gene and its transformation into Capsella bursa-posteris

CRABS CLAW(CRC)是控制拟南芥心皮发育的主要基因之一,属MADS box基因家族中的成员.根据GenBank收录的CRABS CLAW基因的cDNA序列设计引物,通过RT-PCR的方法,从哥伦比亚型拟南芥总RNA中扩增出CRABS CLAW基因的全长cDNA片段,将其克隆到pMD18-T载体中,经测序证明该片段与GenBank报道的序列完全一致.以Ti质粒载体pWM101为基础,构建了由组成型启动子CaMV35S调控的CRC基因的植物表达载体pWM101-CRC,通过根癌农杆菌花序浸渍法转化荠菜,获得了转CRC基因的荠菜植株.CRC基因在荠菜中的组成型表达对荠菜心皮形态和大小都产生了一定影响.

CRABS CLAW（CRC） is a key gene controlling the carpel development ofArabidopsis thaliana, encoding a transcription factor which belongs to MADS family. To clone the gene cDNA a pair of primer was designed and synthesized according to the CRC cDNA sequence. The full length cDNA of the gene was amplified by RT-PCR by using the total RNA as temple that is isolated from the ecotype Colombia seedling ofArabdopsis thaliana. The cDNA was cloned into pMD 18-T vector and be sequenced. DNA sequencing indicated that the cloned fragment is identity to the sequence of CRC cDNA in GenBank and encodes a predicted transcription factor. The cDNA is subsequently cloned into a Ti plasmid pWM101 and be controlled by a consistant promoter 35S of CaMV. The recombinant was transformed into Capsella bursa-posteris which is identical in a heart shaped carple, Transgenic plant obtained and the phenotype of carple varies in both size and shape.