Isolation of the Promoter Region of a Pollen-specific Gene <i>PSG076</i> by Inverse-PCR

CHEN Ling; SU Pei-Pei; TONG Han-Wen; LIU Yi-Ke; ZHU Zhan-Wang; YANG Guang-Xiao; GAO Chun-Bao; HE Guang-Yuan

doi:10.3724/SP.J.1142.2012.30309

Plant Science Journal > 2012 > (3): 309-314. > DOI: 10.3724/SP.J.1142.2012.30309 CSTR: 32231.14.SP.J.1142.2012.30309

CHEN Ling, SU Pei-Pei, TONG Han-Wen, LIU Yi-Ke, ZHU Zhan-Wang, YANG Guang-Xiao, GAO Chun-Bao, HE Guang-Yuan. Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR[J]. Plant Science Journal, 2012, (3): 309-314. DOI: 10.3724/SP.J.1142.2012.30309

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Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR

1.
China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory, The Genetic Engineering International Cooperation Base of Chinese Ministry of Science and Technology, The Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science & Technology (HUST), Wuhan 430074, China
2. Institute of Food Crops, Hubei Academy of Agricultural Sciences, Hubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Wuhan 430064, China

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Received Date: November 20, 2011
Revised Date: March 04, 2012
Published Date: June 29, 2012

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Abstract

Abstract

A pollen-specific gene,PSG076,with unknown function,was isolated from Austrian wheat ‘Ferdinand’.To obtain pollen-specific promoter in wheat genetic engineering,the promoter region of PSG076 gene was isolated by an improved inverse-PCR (IPCR) method.A 1.4 kb promoter sequence was cloned and analyzed by Bioinformatics tools.Some cis-acting elements or enhancers conferred to pollen-specific expression were found,such as AGAAA,GTGA, AGGTCA and AAATGA.This suggested that PSG076 promoter might be a pollen-specific promoter with strong activity.The results also showed that the improved IPCR method was efficient to isolate the flanking unknown sequence,especially in promoter isolation.
- Wheat,
- Pollen-specific promoter,
- Inverse-PCR,
- Sequence analysis

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Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR

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