野生大豆GsGSK1基因的克隆及遗传转化

Cloning and Genetic Transformation of Wild Soybean GsGSK1 Gene into Arabidopsis

以野生大豆为材料,采用同源克隆方法和RT-PCR技术获得一个野生大豆糖原合成酶激酶类基因的全长cDNA(命名为GsGSK1)。该基因的ORF为1 233 bp,推断其编码410氨基酸的多肽。将此基因和与大豆糖原合成酶激酶基因的核苷酸序列比较发现二者有11个核苷酸SNP位点差异,其中,10个为同义突变,1个为非同义突变,同源性达到99.1%。通过构建重组植物表达载体pCAMBIA3300-GsGSK1,将该基因通过农杆菌介导法转入拟南芥,获得了T1抗性苗,为进一步鉴定GsGSK1的基因功能提供了试验基础。

Using homologous cloning and RT-PCR technology,a glycogen synthase kinase family gene,GsGSK1 was isolated from wild soybean.It is 1 233 bp in length with one ORF of 410 amino acids.Compared the nucleotide sequence with GmGSK,these two genes have11 SNPs,in which 10 SNPs are synonymous mutations,and 1 SNP is non-synonymous mutation,with a homology of 99.1%.Then constructed the recombinant plant expression vector pCAMBIA3300-GsGSK1 and transformed it into arabidopsis by agrobacterium mediated method.T1 transgenic plant had been acquired,which will be supplied the basic materials for further identification of gene function in GsGSK1.