拟南芥AtCDPK1基因克隆与转化马铃薯的研究

Clone and Transformation into Potato of AtCDPK1 Gene from Arabidopsis thaliana

以野生拟南芥生态型(Col-0)为材料,利用PCR和DNA重组技术,克隆了拟南芥钙依赖蛋白激酶基因(AtCDPK1)。AtCDPK1基因全长为2 269bp,碱基序列与已知基因At1g18890完全一致。构建AtCDPK1基因的植物表达载体pCHF3-CDPK1,并利用农杆菌介导法将pCHF3-CDPK1转入马铃薯品种‘费乌瑞它’的脱毒试管微型薯中,经筛选与植株再生,共获得50个抗性再生植株。PCR检测显示,有36个植株基因组中含有AtCDPK1基因。RT-PCR分析证实,AtCDPK1基因在这些马铃薯植株的基因组中均能正常转录表达。这一结果为进一步开展AtCDPK1基因功能分析及转基因抗旱马铃薯新品种选育研究奠定了基础。

Using Arabidopsis thaliana（Columbia ecotype）as material,we cloned calcium-dependent protein kinase 1（AtCDPK1）gene with PCR and recombinant DNA technologies.The full length of the gene is 2269 bp,and the sequence of bases was entirely consistent with that of the known gene At1g18890.We constructed the plant expression vector pCHF3-CDPK1 of AtCDPK1gene,and transformed pCHF3-CDPK1 into virus-free miniature of potato cultivar‘Favorita＇using Agrobacterium-mediated method.After plant selection and regeneration,a total of 50 regenerated plants with resistance were obtained successfully.The PCR detection showed that a total of 36 plants contained AtCDPK1 gene,and the RT-PCR analysis proved that the AtCDPK1 gene in transgenic potato plants could conduct normal transcription and expression.This research lays the foundation for further function analysis of AtCDPK1 gene in transgenic potato and breeding new drought-resistant transgenic potato.