摘要
运用已克隆的梅PGIP基因构建植物超表达载体,将PGIP插入带有启动子super-promoter和终止子nos的中间载体P-Super1300+中,酶切鉴定表明,目的基因已正确地插入载体中.用根癌农杆菌介导法将其转入烟草中,共获得25株潮霉素抗性苗,对其中的13株进行PCR检测,有12株扩增出了目的条带;进一步对其中的7株进行Southern杂交检测和RT-PCR检测,发现7个株系均有杂交带出现,且均发生了基因转录,说明已成功获得了能够表达PGIP基因的转基因烟草.
By inserting PGIP in vector P-Superl300^＋ with super-promoter and nos terminator,a plant super-expressed vector of PGIP gene from Prunus mume was constructed successfully and then transferred to tobacco by Agrobacterium-mediated transformation system. Twenty-five resistant plants to hyg were obtained. Among them,thirteen plants were sampled to detect target fragment,and the results showed that the target fragment was detected in twelve plants. Further analysis on seven plants of them showed that they all have southern-blot bands and transcriptional patterns. As a result, transgenic tobacco in which PGIP could be expressed successfully had been obtained.
出处
《西北植物学报》
CAS
CSCD
北大核心
2007年第10期1948-1952,共5页
Acta Botanica Boreali-Occidentalia Sinica
关键词
梅
PGIP基因
表达载体构建
表达载体遗传转化
Prunus mume
PGIP gene
construction of expressed vector
transformation of expressed vector
作者简介
李广平（1974-），女（汉族），博士，讲师，主要从事果树分子生物学方面研究。E—mail：liguangpjng108@sina．com
通讯作者：章镇，博士，教授，主要从事果树学研究。E—mail：zhangzhen_nj@hotmail．com
