Rice Repetitive DNA Sequence RRD3: a Plant Promoter and Its Application to RNA Interference

Jian Zhong^{a, b
, ,},
Hongbin Wang^b,
Dangquan Zhang^{b, c},
Bing Liu^b,
Jinfa Wang^b

a.
National Engineering Center of Cell Products, Amcellgene Co., Ltd., Tianjin 300457, China
b.
State Key Laboratory for Biocontrol and Key Laboratory of Gene Engineering of Ministry of Education, School of Life Sciences of Sun Yet-sun University, Guangzhou 510275, China
c.
Key Lab of Non-Wood Forest Products of Forestry Ministry, Central South University of Forestry and Technology, Changsha 410004, China

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Corresponding author: E-mail address: zhongjianzhj@yahoo.com.cn (Jian Zhong)

摘要

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Abstract: Previously, a moderately repetitive DNA sequence (RRD3) was cloned from rice (Oryza sativa L.) by DNA renaturation kinetics. Sequence analysis revealed several conserved promoter motifs, including four TATA-boxes and a CAAT-box, and promoter activity was shown in Escherichia coli and mammalian expression systems. Here, we inserted the RRD3 fragment into the plant promoter-capture vector, pCAMBIA1391Z, and examined whether the RRD3 fragment has promoter activity in plants. Transgenic tobacco and rice calli both showed β-glucuronidase (GUS) activity, indicating that RRD3 can act as a promoter in both monocot and dicot plants. Based on the promoter characteristic of RRD3, we designed a plant universal binary vector, pCRiRRD3, which is suitable for performing researches on plant RNA interference. This vector has two multiple cloning sites to facilitate sense and antisense cloning of the target sequence, separated by an intron fragment of 200 bp. The efficiency of the vector for gene silencing was assayed by histochemical and quantitative fluorometric GUS assays in transgenic tobacco. These research results suggested that this plant RNAi vector pCRiRRD3 can effectively perform gene silencing researches on both monocot and dicot plants.
- RRD3 /
- promoter /
- GUS histochemical assay /
- quantitative fluorometric GUS assay /
- RNA interference

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参考文献(32)

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