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一种新型组成型玉米启动子在转基因小麦和玉米中的表达特性分析

Characterisation of the expression of a novel constitutive maize promoter in transgenic wheat and maize.

作者信息

Rasco-Gaunt S, Liu D, Li C P, Doherty A, Hagemann K, Riley A, Thompson T, Brunkan C, Mitchell M, Lowe K, Krebbers E, Lazzeri P, Jayne S, Rice D

机构信息

DuPont Wheat Transformation Laboratory, c/o IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, UK.

出版信息

Plant Cell Rep. 2003 Feb;21(6):569-76. doi: 10.1007/s00299-002-0552-y. Epub 2002 Nov 16.

Abstract

A novel constitutive promoter from the maize histone H2Bgene was recently identified. In this study, we characterised H2B promoter activity in both wheat and maize tissues using the gusA reporter gene and two synthetic versions of the pat (phosphinothricin acetyl transferase) selectable marker gene, namely mopat and popat. Analyses of transgenic plants showed that the H2B promoter is able to drive the expression of gusA to strong, constitutive levels in wheat and maize tissues. Using an H2B:mopat construct and phosphinothricin selection, we recovered transgenic wheat plants at efficiencies ranging from 0.3% to 7.4% (mean 1.6%), and the efficiency of selection ranged from 40% to 100% (mean 77.7%). In another application, H2B was combined with the maize Ubi-1 or the maize Adh-1 intron to drive the expression of mopat and popat. Transformation efficiencies with the Ubi-1 intron were between 1.4- to 16-fold greater than with the Adh-1 intron. However, the use of either of the introns was necessary for the recovery of transgenic plants. Mopat gave higher transformation efficiencies and induced higher levels of PAT protein in maize tissues than popat.

摘要

最近鉴定出一种来自玉米组蛋白H2B基因的新型组成型启动子。在本研究中,我们使用gusA报告基因以及两个合成版本的pat(膦丝菌素乙酰转移酶)选择标记基因,即mopat和popat,对小麦和玉米组织中的H2B启动子活性进行了表征。对转基因植物的分析表明,H2B启动子能够驱动gusA在小麦和玉米组织中高水平、组成型表达。使用H2B:mopat构建体和膦丝菌素选择,我们获得转基因小麦植株的效率为0.3%至7.4%(平均1.6%),选择效率为40%至100%(平均77.7%)。在另一项应用中,H2B与玉米Ubi-1或玉米Adh-1内含子结合以驱动mopat和popat的表达。使用Ubi-1内含子的转化效率比使用Adh-1内含子高1.4至16倍。然而,使用任何一个内含子都是获得转基因植株所必需的。在玉米组织中,mopat比popat具有更高的转化效率并诱导更高水平的PAT蛋白。

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