破色期番茄果实均一化cDNA沉默文库的构建和功能基因筛选模型的初步建立

被引：6

作者：

王晓光

马远征

王晓辉

李玲

罗云波

朱本忠

机构：

[1] 中国农业大学食品科学与营养工程学院食品生物技术实验室

来源：

生物化学与生物物理进展 | 2009年 / 08期

关键词：

cDNA文库; 均一化; 功能基因; VIGS;

D O I：

暂无

中图分类号：

S641.2 [番茄（西红柿）];

学科分类号：

摘要：

番茄果实的成熟是由多基因精细调控的一个过程.利用破色期番茄果实,根据复性动力学原理在mRNA水平进行均一化操作使高丰度和低丰度的mRNA丰度接近.然后把均一化之后mRNA反转录得到cDNA,再与基因沉默载体pTRV重组,最后把构建好的载体通过电转化的方法转入到GV3101农杆菌中,从而建立起破色期番茄果实均一化cDNA沉默文库.通过番茄果实中病毒诱导基因沉默技术,对cDNA沉默文库进行初步筛选,从而确定功能基因筛选模型.在模型建立阶段,以番茄红素合成途径相关的PDS基因作为内标基因,在100个混合农杆菌样中,成功筛选到了PDS基因.

引用

页码：1079 / 1083

页数：5

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