DHDPS突变基因作为转基因植物筛选标记的研究

Study on Mutated DHDPS Gene as Selectable Marker of Transgenic Plant

从烟草(Nicotiana tabacum L.cv)中克隆得到赖氨酸合成过程中的关键酶基因二氢吡啶二羧酸合酶(dihydrodipicolinate synthase,DHDPS)基因(dhdps),对其进行体外分子改造,以解除末端产物对它的反馈抑制。将突变基因用做转基因植物的筛选标记转化烟草,用赖氨酸类似物(S-2-氨基乙基-L-半胱氨酸,AEC)作为筛选剂,成功筛选到了PCR阳性植株。Real-time PCR结果表明,PCR阳性植株中dhdps基因的转录水平显著高于对照,但植株表型异常。

The rodipicolinate synthase gene was cloned from Nicotiana tabacum L. cv, which encodes the key enzyme in the synthesis of lysine, and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent, and the detection results were positive by means of PCR and Real-Time PCR, however the phenotype is somewhat abnormal.