论文部分内容阅读
根据拟南芥(Arabidopsisthaliana)、烟草(Nicotianatabacum)EIN3基因保守区序列设计简并引物,采用RT-PCR扩增从番茄(Lycopersiconesculentum )果实中得到一个486bp的同源片段。以此片段作为探针,从番茄果实cDNA文库中分离到一个拟南芥EIN3的同源基因的cDNA克隆,命名为Le-EIL。该克隆含有一个1845bp的开放阅读框,编码615个氨基酸。经同源分析,它与烟草TEIL基因的同源性为79%,与拟南芥EIN3、EIL1、EIL2和EIL3的同源性分别为59%、56%、52%、46%。Le-EIL基因表达与番茄果实的成熟及乙烯生成情况具有较强的相关性。在普通番茄果实的不同成熟时期Le-EIL基因都有表达,并且随着成熟度的增加基因表达有明显的增强,在转色期和粉红期Le-EIL基因的表达最强,在全红期后减弱。在转反义ACS基因的转基因番茄果实中,只有开花后60 d的果实中Le-EIL基因有微弱的表达,在其它成熟时期均没有表达。
Degenerate primers were designed based on the conserved region of Arabidopsishaliana and Nicotianatabacum, and a 486bp homologous fragment was obtained from the fruit of Lycopersicon esculentum by RT-PCR. Using this fragment as a probe, a cDNA clone of a homologue of Arabidopsis thaliana EIN3 was isolated from a tomato fruit cDNA library and named Le-EIL. This clone contains an 1845bp open reading frame encoding 615 amino acids. Homology analysis showed that it shared 79% identity with tobacco TEIL gene and 59%, 56%, 52%, 46% identity with Arabidopsis EIN3, EIL1, EIL2 and EIL3, respectively. Le-EIL gene expression and tomato fruit ripening and ethylene production has a strong correlation. Le-EIL gene was expressed in different ripening stages of common tomato fruit, and the expression of Le-EIL gene was enhanced with the increase of maturity. The expression of Le-EIL gene was the strongest in the chromogenic and pink phases. After weakening. Among the transgenic tomato fruits transformed with antisense ACS gene, only Le-EIL gene was weakly expressed in the fruit at 60 d after flowering, and was not expressed in other mature stages.