Rahman, M. Azizur, Hwang, Gyu-Lin, Razak, Shaharudin Abdul, Sohm, Frederic, Farahmand, Hamid, Smith, Alan, Brooks, Carly and Maclean, Norman (2003) Isolation and characterisation of tilapia β-actin promoter and comparison of its activity with carp β-actin promoter. BBA - Biochimica et Biophysica Acta - Gene Structure and Expression, 1625 (1). pp. 11-18. ISSN 0006-3002 (doi:10.1016/S0167-4781(02)00534-1)

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Abstract

The regulatory sequence including proximal promoter, untranslated exon 1 and intron 1 of the β-actin gene from tilapia (Oreochromis niloticus) has been isolated and spliced to a β-galactosidase reporter gene to test its activity. Comparisons of promoter activity have been carried out with three different constructs: (1) 1.6 kb tilapia β-actin regulatory sequence, (2) 1.5 kb carp β-actin regulatory sequence, and (3) 4.7 kb carp β-actin regulatory sequence. Although the 1.6 kb tilapia β-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 β-actin regulatory sequence by quantitative assay. In comparison with the 1.5 kb carp β-actin regulatory sequence, the 1.6 kb tilapia β-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 β-actin regulatory sequence showed three to four times better activity in blue gill cells than either the 4.7 kb carp β-actin or the 1.5 kb carp β-actin regulatory sequences. The 1.6 kb tilapia β-actin regulatory sequence also drove higher reporter gene activity in somatic cells of tilapia than did the 4.7 kb carp β-actin regulatory sequence following direct injection of constructs into muscle. Therefore, taken together, the data demonstrate that the tilapia β-actin promoter can be used as an efficient regulatory sequence to produce autotransgenic tilapia.

Item Type:	Article
Uncontrolled Keywords:	β-actin gene; Promoter; Tilapia; Transgenic fish
Faculty / School / Research Centre / Research Group:	Faculty of Education, Health & Human Sciences > School of Human Sciences (HUM)
Last Modified:	09 Oct 2021 04:46
URI:	http://gala.gre.ac.uk/id/eprint/14495

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