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Enhanced transgene expression in rice following selection controlled by weak promoters

文献类型： 外文期刊

作者： Zhou, Jie 1 ; Yang, Yong 1 ; Wang, Xuming 1 ; Yu, Feibo 1 ; Yu, Chulang 1 ; Chen, Juan 2 ; Cheng, Ye; Yan, Chenqi; Che 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, MOA Key Lab Plant Protect & Biotechnol, Zhejiang Prov Key Lab Plant Virol, Hangzhou 310021, Zhejiang, Peoples R China

2.Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sust

关键词： tCUP;Promoter activity;Selectable marker;Transgene expression;Copy number;Rice

期刊名称：BMC BIOTECHNOLOGY （ 影响因子：2.563； 五年影响因子：3.292 ）

ISSN： 1472-6750

年卷期： 2013 年 13 卷

页码：

收录情况： SCI

摘要： Background: Techniques that enable high levels of transgene expression in plants are attractive for the commercial production of plant-made recombinant pharmaceutical proteins or other gene transfer related strategies. The conventional way to increase the yield of desired transgenic products is to use strong promoters to control the expression of the transgene. Although many such promoters have been identified and characterized, the increase obtainable from a single promoter is ultimately limited to a certain extent. Results: In this study, we report a method to magnify the effect of a single promoter by using a weak promoter-based selection system in transgenic rice. tCUP1, a fragment derived from the tobacco cryptic promoter (tCUP), was tested for its activity in rice by fusion to both a beta-glucuronidase (GUS) reporter and a hygromycin phosphotransferase (HPT) selectable marker. The tCUP1 promoter allowed the recovery of transformed rice plants and conferred tissue specific expression of the GUS reporter, but was much weaker than the CaMV 35S promoter in driving a selectable marker for growth of resistant calli. However, in the resistant calli and regenerated transgenic plants selected by the use of tCUP1, the constitutive expression of green fluorescent protein (GFP) was dramatically increased as a result of the additive effect of multiple T-DNA insertions. The correlation between attenuated selection by a weak promoter and elevation of copy number and foreign gene expression was confirmed by using another relatively weak promoter from nopaline synthase (Nos). Conclusions: The use of weak promoter derived selectable markers leads to a high T-DNA copy number and then greatly increases the expression of the foreign gene. The method described here provides an effective approach to robustly enhance the expression of heterogenous transgenes through copy number manipulation in rice.

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