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[1]苏军,管其龙,陈子强,等.水稻arf1基因3′-UTR片段的克隆和验证[J].应用与环境生物学报,2019,25(02):414-419.[doi:10.19675/j.cnki.1006-687x.2018.05023]
 SU Jun**,GUAN Qilong,CHEN Ziqian,et al.Cloning and validation of 3′-UTR fragments of rice arf1 gene[J].Chinese Journal of Applied & Environmental Biology,2019,25(02):414-419.[doi:10.19675/j.cnki.1006-687x.2018.05023]
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水稻arf1基因3′-UTR片段的克隆和验证()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年02期
页码:
414-419
栏目:
研究论文
出版日期:
2019-04-25

文章信息/Info

Title:
Cloning and validation of 3′-UTR fragments of rice arf1 gene
作者:
苏军管其龙陈子强陈在杰
福建省农业科学院生物技术研究所,福建省农业遗传工程重点实验室 福州 350003
Author(s):
SU Jun** GUAN Qilong CHEN Ziqian CHEN Zaijie
Fujian Provincial Key Laboratory of Genetic Engineering for Agriculture, Biotechnology Institute of Fujian Academy of Agricultural Sciences, Fuzhou 350003, China
关键词:
水稻3′-UTR终止子克隆表达
Keywords:
rice 3′-UTR terminator clone expression
分类号:
Q943.2
DOI:
10.19675/j.cnki.1006-687x.2018.05023
摘要:
终止子是一段位于基因编码之后的序列,作为一种调控信号调控DNA的转录终止和RNA的释放,在基因工程技术应用中通常被构建在目的基因下游,用以调控基因的转录和表达. 现有常用的终止子很少,克隆和验证新的终止子是植物基因工程技术发展的需要. 通过生物信息学分析和Realtime-PCR表达验证,筛选出候选基因腺苷酸核糖基化作用因子(Similar to ADP-ribosylation factor1,arf1)基因,克隆了水稻arf1基因3′-UTR. 结果显示,所克隆3′-UTR片段含有8个多聚信号元件和4个UE片段. 将3′-UTR与gus基因融合后分别与玉米泛素启动子Ubiquitin和花椰菜花叶病毒(CaMV)启动子35S连接构建植物表达载体验证3′-UTR调控表达效果. 烟草瞬时表达显示3′-UTR融合的gus?基因在35S和Ubi启动子驱动下,在烟草叶片中能正常表达;3′-UTR调控下的gus基因在水稻根、茎、叶、花和种子中均可稳定表达,且表达量与T-Nos终止子相当. 本研究表明所克隆的3′-UTR可替代T-nos终止子,可为植物基因工程技术的应用提供新的调控元件. (图5 表3 参29)
Abstract:
A terminator is a sequence that is usually located at the end of a gene, and is a regulatory signal to block the transcription of DNA to RNA and release the transcript. At present, there are limited commonly used terminators; therefore, the cloning and validation of new terminators is required to develop genetically modified crops. Here, we cloned a 3′-UTR sequence of a candidate gene similar to ADP-ribosylation factor, arf1, and analyzed the control element of the sequence. The sequence of arf1 3′-UTR contained eight polyA sites and four U-rich elements. To confirm that the 3′-UTR of the arf1 gene is an important component for the regulation of transcript stability, two reporter gene constructs were made by cloning the arf1 gene 3′-UTR downstream of the bacterial b-glucuronidase gene (uidA) under the control of the CaMV 35S and Ubi promoters. An agrobacterium-mediated transient gene expression test showed that the 3′-UTR–fusion gus gene driven by the 35S or Ubi promoters can be normally expressed in tobacco leaves. The gus gene expression regulated by 3′-UTR was stable in the rice roots, stems, leaves, flowers, and seeds, and the expression level was comparable to that of T-Nos. The study showed that the clone 3′-UTR can replace the T-Nos terminator, and the results provide new regulatory elements for the application of plant genetic engineering technology.

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更新日期/Last Update: 2019-04-25