Author:
Al-Robaiy Samiya,Eschrich Klaus
Abstract
AbstractThe 1282 bp cDNA of an isoenzyme of fructose-1,6- bisphosphatase was cloned from rat muscle. It shows 70% positional identity to the cDNA of rat liver fructose- 1,6-bisphosphatase and is clearly the product of a gene different from that coding for the liver enzyme. After cloning of the coding region of the rat muscle fructose-1,6-bisphosphatase cDNA in an expression vector, the recombinant enzyme could be detected inE. colicell-free extracts by activity determination and Western blotting. Overexpressed fructose-1,6-bisphosphatase was found to be allosterically inhibited by AMP comparably to the enzyme isolated from rat muscle. Analysis of steady-state mRNA levels of various rat tissues with reverse-transcriptase polymerase chain reaction (RT-PCR) and Northern blotting revealed one or the two fructose-1,6-bisphosphatase isoenzyme mRNAs in most tissues tested with significant quantitative differences. Quantitative PCR using a homologous competitor showed that 1 μg of total RNA of rat muscle contains 1.7 × 106molecules of rat muscle fructose-1,6-bisphosphatase mRNA. 3 × 104copies of this message were found per μg total RNA of heart and kidney, respectively.
Subject
Clinical Biochemistry,Molecular Biology,Biochemistry
Cited by
32 articles.
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