Belouski E, Watson D E, Bennett G N
Department of Biochemistry and Cell Biology-MS 140, 6100 Main St., Rice University, Houston, TX 77005-1892, USA.
Curr Microbiol. 1998 Jul;37(1):17-22. doi: 10.1007/s002849900330.
The pfk gene encoding phosphofructokinase (Pfk) from the anaerobic bacterium Clostridium acetobutylicum ATCC 824 was cloned and sequenced. The gene was identified in a plasmid library by complementation of an E. coli pfk mutant and by the ability to amplify a fragment by PCR using primers based on homologous regions of Pfk from other microorganisms. Nucleotide sequence analysis revealed a coding region for a 319-aa protein homologous to Pfks from other organisms. Enzyme assay and ability to complement the growth defects of E. coli pfk mutants confirmed the expression of the clostridial pfk gene. The pyruvate kinase (pyk) gene was identified adjacent to pfk. Such an arrangement for the genes encoding key regulators of glycolytic flux had not yet been described in a strict anaerobe. This gene arrangement has been found in other Gram-positive organisms, but not in Gram-negative organisms.
对来自厌氧细菌丙酮丁醇梭菌ATCC 824的编码磷酸果糖激酶（Pfk）的pfk基因进行了克隆和测序。通过对大肠杆菌pfk突变体的互补作用以及使用基于其他微生物Pfk同源区域的引物通过PCR扩增片段的能力，在质粒文库中鉴定出该基因。核苷酸序列分析揭示了一个编码319个氨基酸蛋白质的编码区，该蛋白质与其他生物体的Pfk同源。酶活性测定以及互补大肠杆菌pfk突变体生长缺陷的能力证实了梭菌pfk基因的表达。丙酮酸激酶（pyk）基因被鉴定为与pfk相邻。这种编码糖酵解通量关键调节因子的基因排列在严格厌氧菌中尚未见报道。这种基因排列已在其他革兰氏阳性生物中发现，但在革兰氏阴性生物中未发现。
