alfAFP和spCEMA融合基因表达载体的构建及其对小麦的遗传转化

Co-transformation of wheat with fusion genes alfAFP and spCEMA using biolistics

以小麦品种西农1376为受体,利用Overlapping PCR的方式构建了防御素基因alfAFP和抗菌肽基因spCEMA融合基因的表达载体,与pAHC20共转化幼胚愈伤组织,获得转基因再生T0代植株269株,春化后移栽成活188株。对叶片基因组DNA进行目的基因和bar基因PCR扩增表明其中7株是共整合植株,共整合频率为0.1%,初步证明融合基因已成功通过共转化方式导入受体小麦品种中。影响共整合率的因素分析表明,在使用除草剂PPT对bar基因进行筛选的过程中,筛选时间的缩短并不会明显降低筛选的效率。

Transgenic wheat variety XiNong 1376 containing the plant defensin gene alfAFP and antimicrobial peptide gene spCEMA fusion gene were obtained using cotransformation with bar as a selective marker via biolistics.In total,269 plantlets were generated through screening,and 188 of which were alive after vernalization.Leaf genome DNA PCR array showed that 7 co-transformed plants were obtained,and the frequency of co-integration was 0.1%.Studies also illustrated that the shortened time of the bar gene screening process did not significantly reduce the efficiency of PPT screening.