半胱氨酸蛋白酶基因表达载体的构建及转基因苜蓿的初步研究

Construction of plant expression vectors containing MsCP1 genes and preliminary study of transgenic alfalfa

【目的】为培育高抗逆的作物品种提供科学依据。【方法】以耐盐苜蓿"中苜1号"为外植体,采用根癌农杆菌(Agrobacterium tumefaciens)介导的叶盘法转化苜蓿。【结果】成功构建了含有半胱氨酸蛋白酶基因MsCP1和报告基因GUS的植物表达载体pCAMBLA1391-CP-GUS,共获得47株抗性再生植株。经PCR和RT-PCR检测,表明外源基因已成功整合到苜蓿基因组中,并在转录水平得到表达。【结论】初步获得了高抗逆的转基因苜蓿新品系。

[Objective]This study was done in order to give new insights into the breeding of high stress- resistant crop cultivars. [Method] The genetic transformation of alfalfa cultivars ＂Zhongmu No. 1＂ mediated by Agrobacterirn turnefaciens was studied by using the transient expression of GUS gene. IRe- suit] A new plant expression vector pCAMBLA1391-CP-GUS, containing cysteine protease gene MsCP1 and GUS gene,was successfully constructed through sequentia.1 restriction digests and ligations recombina- tion. 47 transformation plants were obtained. PCR and RT-PCR analysis showed that the foreign MsCP1 and GUS gene were inserted into the genome of these plants and expressed in mRNA level. [Conclusion] New strains of transgenic alfalfa with high stress-resistance were primarily gained.