PUBLICATION
Isolation and characterisation of tilapia beta-actin promoter and comparison of its activity with carp beta-actin promoter
- Authors
- Hwang, G.L., Azizur Rahman, M., Abdul Razak, S., Sohm, F., Farahmand, H., Smith, A., Brooks, C., and Maclean, N.
- ID
- ZDB-PUB-090210-18
- Date
- 2003
- Source
- BBA Gene Structure and Expression 1625(1): 11-18 (Journal)
- Registered Authors
- Maclean, Norman
- Keywords
- β-actin gene, Promoter, Tilapia, Transgenic fish
- MeSH Terms
-
- Actins/genetics*
- Animals
- Animals, Genetically Modified
- Carps/embryology
- Carps/genetics*
- Microinjections
- Promoter Regions, Genetic*
- Sequence Analysis, DNA
- Tilapia/embryology
- Tilapia/genetics*
- Transgenes
- PubMed
- 12527420 Full text @ BBA Gene Structure and Expression
Citation
Hwang, G.L., Azizur Rahman, M., Abdul Razak, S., Sohm, F., Farahmand, H., Smith, A., Brooks, C., and Maclean, N. (2003) Isolation and characterisation of tilapia beta-actin promoter and comparison of its activity with carp beta-actin promoter. BBA Gene Structure and Expression. 1625(1):11-18.
Abstract
The regulatory sequence including proximal promoter, untranslated exon 1 and intron 1 of the beta-actin gene from tilapia (Oreochromis niloticus) has been isolated and spliced to a beta-galactosidase reporter gene to test its activity. Comparisons of promoter activity have been carried out with three different constructs: (1) 1.6 kb tilapia beta-actin regulatory sequence, (2) 1.5 kb carp beta-actin regulatory sequence, and (3) 4.7 kb carp beta-actin regulatory sequence. Although the 1.6 kb tilapia beta-actin regulatory sequence gave slightly different expression patterns in tilapia embryos assayed by in situ X-gal staining, no difference was observed in expression level when the tilapia sequence was compared with the 4.7 kb carp beta-actin regulatory sequence by quantitative assay. In comparison with the 1.5 kb carp beta-actin regulatory sequence, the 1.6 kb tilapia beta-actin regulatory sequence gave higher expression levels in tilapia embryos, while a reverse result was observed in zebrafish embryos. In cell transfection experiments, the 1.6 kb tilapia beta-actin regulatory sequence showed three to four times better activity in blue gill cells than either the 4.7 kb carp beta-actin or the 1.5 kb carp beta-actin regulatory sequences. The 1.6 kb tilapia beta-actin regulatory sequence also drove higher reporter gene activity in somatic cells of tilapia than did the 4.7 kb carp beta-actin regulatory sequence following direct injection of constructs into muscle. Therefore, taken together, the data demonstrate that the tilapia beta-actin promoter can be used as an efficient regulatory sequence to produce autotransgenic tilapia.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping