Introduction and expression of the bacterial glyoxylate cycle genes in transgenic mice
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The glyoxylate cycle, catalysed by two unique enzymes: isocitrate lyase (ICL; EC 188.8.131.52) and malate synthase (MS; EC 184.108.40.206), is necessary for the net conversion of acetate into glucose. This metabolic pathway operates in microorganisms, higher plants and nematodes. Two bacterial genes, encoding ICL and MS, were modified in order to introduce them into the mouse germ line. The ovine metallothionein-Ia (MT-Ia) promoter-aceB gene-ovine growth hormone (GH) gene (3′ GH sequence) construct was fused to the ovine MT-Ia promoter-aceA gene-ovine GH gene (3′ GH sequence). Therefore, in this single DNA sequence, bothaceA andaceB are under independent MT-Ia promoter control and can be induced by zinc. Transgenic mice were generated by pronuclear microinjection of theaceB-aceA gene construct. We now report the establishment of four mouse lines carying these two transgenes. Studies on the progeny of these lines indicate that one line (No. 91) is expressing both genes at the mRNA and enzyme levels in the liver and intestine, whereas another line (No. 66) has a much lower expression. Both enzyme activities were detected in the liver and intestine at levels up to 25% of those measured in fully derepressedEscherichia coli cells.
Keywordsglyoxylate cycle isocitrate lysae malate synthase microinjection transgenic mice expression
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