【摘 要】
：
本研究在前人对番木瓜β-GAL多基因家族研究的基础上,采用IPCR技术进行了三次步移,获得了β-GAL同源基因5’端和启动子区,并对其进行了功能鉴定,为将来采用启动子区RNAi技术
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本研究在前人对番木瓜β-GAL多基因家族研究的基础上,采用IPCR技术进行了三次步移,获得了β-GAL同源基因5’端和启动子区,并对其进行了功能鉴定,为将来采用启动子区RNAi技术研究β-GAL多基因家族的各个成员在番木瓜果实软化中的作用大小,延长番木瓜果实货价期奠定了基础。具体结论如下:1、完成了番木瓜β-GAL同源基因5’端和启动子的克隆和序列分析。利用IPCR技术克隆出番木瓜β-GAL同源基因5’端和启动子,并登陆GenBank,序列号为EF694758。利用在线软件Neural Netwok Promoter Prediction对基础启动子区和转录起始位点进行预测,结果发现在195-245bp,552-602bp,560-610bp和591-641bp的位置可能存在基础启动子序列。并采用启动子预测软件Plant CARE和PLACE对该序列进行分析,分析发现,其中具有大多数高等植物启动子的保守元件,并具有一些和果树成熟有关的调控元件,如ABA反应元件。2、完成了启动子功能的初步检测。构建了启动子缺失表达载体,通过农杆菌介导,进行了GUS瞬时检测,初步说明β-GAL同源基因启动子在番木瓜果实中具有启动功能。通过染色时间和染色程度推测:在603bp和724bp之间可能存在抑制启动的元件。
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