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通过分子生物学技术把酸性蛋白酶pepB基因转入受体玉米自交系基因组中,培育转酸性蛋白酶pepB基因玉米新品系。以耐盐基因badh作为筛选标记性基因,经过抗性筛选,对T1、T2、T3代转基因植株进行PCR检测,得到14个T1代转基因阳性植株,32个T2代转基因阳性植株和27个T3代转基因阳性植株。Southern杂交结果表明,外源酸性蛋白酶基因已经整合进玉米基因组中。RT-PCR结果表明,酸性蛋白酶基因在受体玉米中获得表达,获得外源酸性蛋白酶pepB基因遗传表达的T3代转基因玉米株系。通过对T2、T3代转基因玉米各品系农艺性状的分析结果表明,转基因玉米各品系在株高、茎粗、穗长等农艺性状上与受体亲本没有差异,但生育期均有不同程度的缩短。
The pepB gene of acid protease was transferred into the recipient maize inbred line genome by molecular biology technique to cultivate a new maize line of the acid protease pepB gene. Using salt-tolerant gene badh as screening marker gene, PCR was carried out on T1, T2 and T3 generation transgenic plants after resistance screening, and 14 T1-transgenic plants, 32 T2-transgenic plants and 27 T3 plants Transgenic positive plants. Southern hybridization results show that the exogenous acid protease gene has been integrated into the genome of corn. The results of RT-PCR showed that the acid protease gene was expressed in recipient maize and the T3 generation transgenic maize strain with the genetic expression of exogenous acid protease pepB gene was obtained. Through analyzing the agronomic traits of T2 and T3 generations of transgenic maize, the results showed that the agronomic traits of transgenic maize did not differ from the recipient parents in agronomic traits such as plant height, stem diameter and spike length, but all of them had different degrees of shortening .