【摘要】 从番茄果实中提取总RNA,根据GeneBank中LeETR1序列,设计合成特异性引物,利用RT-PCR技术克隆了LeETR1基因3’端非编码区313 bp的cDNA,经酶切图谱和序列分析鉴定无误后,反向插入到植物表达载体pPZP111A中,构建了表达LeETR1反义RNA的双元载体。经农杆菌途径转化番茄品种B1后,通过PCR检测从抗卡那霉素再生植株中筛选到13株阳性植株,Southem blot杂交确证反义基因已经整合到番茄染色体中。对果实乙烯释放的测定结果表明,转基因番茄乙烯释放高峰的出现比对照果实推迟10天,番茄红素的合成受到显著抑制,果实不能形成正常的红色。推测LeETR1和番茄的成熟有着密切的关系。更多还原

【Abstract】 Total RNA was isolated from the pink tomato fruits. By using the specific primers designed from the LeETRl cDNA, a 313bp fragment on 3’ encoding region was produced by reverse transcription poly-merase chain reaction(RT-PCR). The cloned LeETR1 gene was then inserted into a binary vector pPZP111A. in an inverted orientation between the CaM 35S promoter and the Nos 3’ termination sequence. Transgenic tomato plants were obtained by Agrobacterium-mediated transformation of cotyledons. PCR detection and Southern blot analysis confirmed the integration of antisence LeETR1 gene in tomato genome. Ethylene production peak of transgenic fruits appeared 10d after control. After 30 days in normal room, transgenic tomatos evevtually developed an orange colour but never turned red. These datas suggest that LeETRl was closely correlated with the ripening of tomato fruits.更多还原