Functional Analysis of Elements Affecting Expression of the β-Actin Gene of Carp

Z. Liu, B. Moav, A. J. Faras, K. S. Guise, A. R. Kapuscinski, P. B. Hackett

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82 Scopus citations

Abstract

Regulatory regions of the β-actin gene of the common carp (Cyprinus carpio) have been examined by linking upstream, 5′-flanking sequences and regions of the first intron to a bacterial chloramphenicol acetyltransferase (CAT) reporter gene. By analysis of the mRNA products and encoded CAT activity, we have identified four putative regions that influence expression: (i) a negative regulatory region 2,300 to 1,100 base pairs (bp) ahead of the gene; (ii) a proximal promoter element, containing the highly conserved CCAAT, CC(A/T)6GG, and TATA boxes that is within the first 204 bp upstream of the initiation site; (iii) a negative element of 426 bp in the 5′ the first intron; and (iv) a positive 304-bp element near the end of the first intron that contains highly conserved sequences found in all characterized β-actin genes. The positive intron element is not a classical enhancer; it is position and orientation dependent, as has been observed in other housekeeping genes in vertebrates. Depending on the elements joined together, CAT gene expression can be modulated more than 500-fold in transfected mouse cells.

Original languageEnglish (US)
Pages (from-to)3432-3440
Number of pages9
JournalMolecular and cellular biology
Volume10
Issue number7
StatePublished - Jul 1990

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