马铃薯蛋白激酶基因StPki的遗传转化及表达分析

Genetic Transformation of Diploid Potato and Expression Suppression of Its Protein Kinase Gene StPki

以高抗青枯病二倍体马铃薯基因型ED13为材料,克隆了蛋白激酶基因StPki。以StPki基因特异区段为靶标,成功构建了该基因的RNA干扰植物表达载体pCHF1-StPki。利用重组农杆菌株LBA4404(pCHF1-StPki)感染转化ED13茎段外植体,获得了抗庆大霉素的再生植株。利用CaMV35S启动子特异引物对再生植株进行PCR检测,结果表明获得了转基因植株。利用StPki基因的特异引物对转基因植株进行半定量RT-PCR分析,结果显示该基因的转录受到了抑制。马铃薯抗病基因型ED13已被成功转化,且表现出了对StPki基因的RNA干扰活性。

Protein kinase gene StPki was cloned from bacterial wilt-resistant diploid potato ED13,and its RNAi plant expression vector was constructed. The expression vector pCHF1-StPki was introduced into Agrobacterium strain LBA4404 and the recombinant strain LBA4404 （ pCHF1-StPki） was used to transform ED13 stem segment. In the presence of CaMV35S promoter-specific primers,Gen-resistant regeneration plants were detected by PCR and transgenic potato plant producing 500 bp amplification product were obtained. Semi-quantitative RT-PCR analysis indicated that the transcription of StPki gene had beed significantly restrained in the transgenic ED13 plant.