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Isolation and primary function detection of Nile tilapia (Oreochromis niloticus) β-actin promoter

Published online by Cambridge University Press:  01 October 2008

Yu Er-Meng ,
Ye Xing ,
Wang Hai-Ying ,
Bai Jun-Jie ,
Xia Shi-Ling ,
Lao Hai-Hua  and
Lu Mai-Xin
Yu Er-Meng
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China College of Aqua-life Science and Technology, Shanghai Ocean University, Shanghai 200090, China
Ye Xing*
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
Wang Hai-Ying
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
Bai Jun-Jie
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
Xia Shi-Ling
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
Lao Hai-Hua
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
Lu Mai-Xin
Affiliation:
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

A 5′-flanking region and partial open reading frame (ORF) of the β-actin gene (GenBank accession No. EF026001) of Nile tilapia (Oreochromis niloticus) was cloned by polymerase chain reaction (PCR) amplification. The segment included a 1643 bp regulatory sequence and a 90 bp partial ORF which encoded a 30-amino-acid peptide. The regulatory sequence comprised a 108 bp 5′ proximal promoter, the first untranslated exon and the first intron of the β-actin gene. The proximal promoter region contained elements that were critical for transcription activity, including a CCAAT, TATA and CArG box located at –92, –29 and –62 bp upstream of the transcription initiation site, respectively. The regulatory sequence was inserted into the promoterless pDsRed2-1 vector to construct the expressing vector pNA-DsRed. The linearized pNA-DsRed was microinjected into the fertilized eggs of Tanichthys albonubes. The expression of the DsRed2 gene in transgenic fish could be observed under the microfluoroscope and anatomical lens. The results showed that the β-actin gene promoter possessed effective transcription activity.

Keywords

Nile tilapiaβ-actin gene promoterred fluorescent protein (DsRed2)transgenemicroinjection
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 2 , August 2008 , pp. 141 - 146
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(2): 242–247

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