alfAFP和spCEMA融合基因表达载体的构建及其对小麦的遗传转化

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。

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

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.