Croxen R, Goosey M W, Keon J P, Hargreaves J A
Shell Research Ltd, Sittingbourne Research Centre, Kent, UK.
Microbiology (Reading). 1994 Sep;140 ( Pt 9):2363-70. doi: 10.1099/13500872-140-9-2363.
A gene encoding 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase was isolated from the maize fungal pathogen Ustilago maydis. This was accomplished by identifying cDNA and genomic clones that hybridized to an internal fragment of the gene, amplified from U. maydis genomic DNA by PCR. The nature of the gene was determined by nucleotide sequence analysis, and by comparing the derived amino acid sequence of the gene with HMG-CoA reductases from yeast, and from other organisms. The hydrophobic nature of the N-terminal region of the deduced protein sequence also supported the view that this gene encoded HMG-CoA reductase. A C-terminal-truncated fragment of the U. maydis HMG-CoA reductase gene was shown to be expressed in Escherichia coli in a catalytically active form. The expressed protein was also shown to be sensitive to an inhibitor of mammalian HMG-CoA reductase activity.
从玉米真菌病原体玉米黑粉菌（Ustilago maydis）中分离出了一个编码3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶的基因。这是通过鉴定与该基因内部片段杂交的cDNA和基因组克隆来实现的，该内部片段是通过PCR从玉米黑粉菌基因组DNA中扩增得到的。通过核苷酸序列分析，并将该基因推导的氨基酸序列与来自酵母及其他生物体的HMG-CoA还原酶进行比较，确定了该基因的性质。推导的蛋白质序列N端区域的疏水性也支持了该基因编码HMG-CoA还原酶的观点。玉米黑粉菌HMG-CoA还原酶基因的C端截短片段在大肠杆菌中以催化活性形式表达。所表达的蛋白质也显示对哺乳动物HMG-CoA还原酶活性抑制剂敏感。
