Schaap P J, Müller Y, Sonnenberg A S, van Griensven L J, Visser J
Section Molecular Genetics of Industrial Microorganisms, Wageningen Agriculturál University, The Netherlands.
Appl Environ Microbiol. 1997 Jan;63(1):57-62. doi: 10.1128/aem.63.1.57-62.1997.
A fortuitously cloned 3'-truncated cDNA encoding the Agaricus bisporus delta 1-pyrroline-5-carboxylate dehydrogenase was used to characterize the complete gene. The gene would encode a cytosolic polypeptide of 546 amino acids, and the basidiomycetous gene was evenly expressed in various parts of the mushroom except for the gills. No expression was detected in compost-grown mycelium. The steady-state mRNA level of the gene in the vegetative phase was determined on simple synthetic media and was two- to threefold higher with ammonium or proline as the sole nitrogen source compared to glutamate as the sole nitrogen source. Moreover, the steady-state mRNA level was not markedly influenced by addition of ammonium phosphate to proline- or glutamate-utilizing cultures. The results suggest that ammonium and the amino acids proline and glutamate are equally preferred nitrogen sources in this organism and are consistent with previous observations of H. M Kalisz, D.A. Wood, and D. Moore (Trans. Br. Mycol. Soc. 88:221-227, 1987) that A. bisporus continues to degrade protein and secrete ammonium even if ammonium and glucose are present in the culture medium.
一个偶然克隆得到的编码双孢蘑菇Δ1-吡咯啉-5-羧酸脱氢酶的3'-截短cDNA被用于鉴定完整基因。该基因编码一个由546个氨基酸组成的胞质多肽,担子菌基因在蘑菇的各个部位均匀表达,但菌褶除外。在堆肥培养的菌丝体中未检测到表达。在简单合成培养基上测定了该基因在营养阶段的稳态mRNA水平,以铵或脯氨酸作为唯一氮源时,其水平比以谷氨酸作为唯一氮源时高两到三倍。此外,向利用脯氨酸或谷氨酸的培养物中添加磷酸铵对稳态mRNA水平没有显著影响。结果表明,铵以及氨基酸脯氨酸和谷氨酸是该生物体同样优先选择的氮源,这与H.M.卡利什、D.A.伍德和D.摩尔之前的观察结果(《英国真菌学会会刊》88:221 - 227, 1987)一致,即即使培养基中存在铵和葡萄糖,双孢蘑菇仍会继续降解蛋白质并分泌铵。