Hostname: page-component-788cddb947-jbkpb Total loading time: 0 Render date: 2024-10-15T03:11:39.363Z Has data issue: false hasContentIssue false

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
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: gzyexing@163.com

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.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

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

References

Alam, MS, Lavender, FL, Iyengar, A, et al. (1996) Comparison of the activity of carp and rat β-actin gene regulatory sequences in tilapia and rainbow trout embryos. Molecular Reproduction and Development 45: 117122.3.0.CO;2-W>CrossRefGoogle ScholarPubMed
Depontizilli, L, Seilertuyns, A and Paterson, BM (1988) A 40-base-pair sequence in the 3′ end of the β-actin gene regulates β-actin mRNA transcription during myogenesis. Proceedings of the National Academy Sciences of the USA 85: 13891393.CrossRefGoogle ScholarPubMed
Feng, H, Cheng, J, Luo, J, Liu, SJ and Liu, Y (2006) Cloning of blank carp β-actin gene and primarily detecting the function of its promoter region. Acta Genetica Sinica 33(2): 133140.CrossRefGoogle Scholar
Frederickson, RM, Micheau, MR, Iwamoto, A and Miyamoto, NG (1989) 5′ flanking and first intron sequences of the human β-actin gene required for efficient promoter activity. Nucleic Acids Research 17(1): 254270.CrossRefGoogle ScholarPubMed
Hamada, K, Tamaki, K, Sasado, T, et al. (1998) Usefulness of the medaka β-actin promoter investigated using a mutant GFP reporter gene in transgenic medaka (Oryzias latipes). Molecular Marine Biology Biotechnology 7: 173180.Google ScholarPubMed
Hanley, S, Smith, TJ, Muller, F, et al. (1998) Isolation and functional analysis of the histone H3 promoter from Atlantic salmon (Salmo salar L.). Molecular Marine Biology Biotechnology 7: 165172.Google ScholarPubMed
Higashijima, S, Okamoto, H, Ueno, N, Hotta, Y and Eguchi, G (1997) High frequency generation of transgenic zebrafish which reliably express GFP in whole muscles or the whole body by using promoters of zebrafish origin. Developmental Biology 192: 289299.CrossRefGoogle ScholarPubMed
Hwang, GL, Rahman, MA, Razak, SA, et al. (2003) Isolation and characterisation of tilapia β-actin promoter and comparison of its activity with carp β-actin promoter. Biochimica et Biophysica Acta 1625: 1118.CrossRefGoogle ScholarPubMed
Jian, Q, Bai, JJ, Ye, X, Xia, SL, Liang, XF and Luo, JR (2004) Cloning of Mylz2 promoter and generation of green fluorescence transgenic zebrafish. Journal of Fishery Sciences of China 11(5): 391395.Google Scholar
Kozak, M (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44(2): 283292.CrossRefGoogle ScholarPubMed
Kozak, M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Research 15(20): 81258148.CrossRefGoogle ScholarPubMed
Liu, ZJ, Moav, B, Faras, AJ, Guise, KS, Kapuscinski, AR and Hackett, PB (1990a) Functional analysis of elements affecting expression of the β-actin gene of carp. Molecular and Cellular Biology 10: 34323440.Google ScholarPubMed
Liu, ZJ, Moav, B, Faras, AJ, Guise, KS, Kapuscinski, AR and Hackett, PB (1990b) Development of expression vectors for transgenic fish. Bio/Technology 8: 12681272.Google ScholarPubMed
Moav, B, Liu, Z, Caldovic, LD, Gross, ML, Faras, AJ and Hackett, PB (1993) Regulation of expression of transgenes in developing fish. Transgenic Research 2: 153161.CrossRefGoogle ScholarPubMed
Noh, JK, Cho, KN, Han, EH, et al. (2003) Genomic cloning of mud loach Misgurnus mizolepis (Cypriniformes, Cobitidae) β-actin gene and usefulness of its promoter region for fish transgenesis. Marine Biotechnology 5: 244252.CrossRefGoogle ScholarPubMed
Ponte, P, Ng, S, Engel, J, Gunning, P and Kedes, L (1984) Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human β-actin cDNA. Nucleic Acids Research 12: 16871696.CrossRefGoogle ScholarPubMed
Quitschke, WW, Lin, ZY, Depontizilli, L and Paterson, BM (1989) The β-actin promoter: high levels of transcription depend upon a CCAAT binding factor. Journal of Biological Chemistry 264(10): 95399546.CrossRefGoogle ScholarPubMed
Seiler-Tuyns, A, Eldridge, JD and Paterson, BM (1984) Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells. Proceedings of the National Academy Sciences of the USA 81: 29802984.CrossRefGoogle ScholarPubMed
Takagi, S, Sasado, T, Tamiya, G, et al. (1994) An efficient expression vector for transgenic medaka construction. Molecular Marine Biology Biotechnology 3: 192199.Google ScholarPubMed
Wang, HY (2007) Molecular cloning of two different lengths of β-actin gene promoters of Tanichthys albonubes and comparison of their activity. Degree Paper, Shanghai Fisheries University.Google Scholar
Williams, DW, Muller, F, Lavender, FL, Orban, L and Maclean, N (1996) High transgene activity in the yolk syncytial layer affects quantitative transient expression assays in zebrafish (Danio rerio) embryos. Transgenic Research 5: 433442.CrossRefGoogle ScholarPubMed
Ye, X, Wang, HY, Qian, Q, Bai, JJ and Lao, HH (2007) Application research of transgenic aquatic animals. Journal of Guangdong Ocean University 27(1): 95100.Google Scholar
Zhu, XP, Xia, SL, Zhang, Y and Liu, JZ (1997) Preliminary study on transgenic mud carp of antifreeze protein gene. Journal of Fishery Sciences of China 4(2): 7980.Google Scholar