摘要
为了实现外源基因在番茄果实中的高效和特异表达,克隆了番茄果实特异基因多聚半乳糖醛酸酶基因(Poly-galacturonase,PG)的启动子。以中蔬四号番茄为材料,建立并优化了以子叶为外植体的番茄高效再生和遗传转化体系;以GUS为报告基因,构建PG GUS植物表达载体,转化番茄。结果表明,在1.0 mg/L ZT的MS分化培养中,番茄子叶的发芽率最高,芽的诱导率高达91%,且发生畸态芽和褐化的外植体最少;通过抗生素浓度对农杆菌的抑制效果试验发现,当头孢霉素的浓度为200 mg/L时,抑制农杆菌的效果最好;成功克隆了番茄PG启动子,将PG启动子驱动的GUS基因转入番茄,对转基因后代果实的GUS染色表明,PG启动子驱动的外源基因在果实中特异表达。
In order to obtain fruit-specific and highly expressed foreign gene in transgenic tomato,we cloned the promoter of Poly-galacturonase（PG）.The tomato regeneration and transformation system were established using ＂zhongshu 4＂.PG promoter： GUS construct was made and introduced into tomato through Agrobacterium mediated transformation.Results showed that the best callus initiation and shoot induction medium was MS ＋ 1.0 mg/L ZT.The shoots induction rate reached 91% with low portion of abnormal shoots.200 mg/L of cefotaxime could effectively inhibit the growth of Agrobacteria.Histochemical analysis showed that GUS was specifically expressed in transgenic tomato fruit.
出处
《生物技术通报》
CAS
CSCD
北大核心
2012年第1期74-78,共5页
Biotechnology Bulletin
基金
山东省中青年科学家奖励基金项目(2006BS06008)
山东省自然科学基金项目(ZR2010CQ008)
山东省"泰山学者"基金项目
