Functional analysis of elements affecting expression of the beta-actin gene of carp.
Regulatory regions of the beta-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' region of 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 beta-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.
Liu
Z J
ZJ
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108.
Moav
B
B
Faras
A J
AJ
Guise
K S
KS
Kapuscinski
A R
AR
Hackett
P B
PB
D000199:Actins / Q000235:genetics*; D000818:Animals; D002347:Carps; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005796:Genes; D005822:Genetic Vectors; D007438:Introns; D007739:L Cells / Q000378:metabolism; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014162:Transfection
D000199:Actins