Templeton M D, Greenwood D R, Beever R E
Molecular Genetics Group, Horticulture and Food Research Institute of New Zealand.
Exp Mycol. 1995 Jun;19(2):166-9. doi: 10.1006/emyc.1995.1020.
Proteins from conidial rodlet preparations of Neurospora crassa were solubilized in trifluoroacetic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of solubilized rodlets revealed a predominant protein of approximately 7 kDa. This protein was absent from preparations of N. crassa cultures carrying the eas mutation. The protein was purified by reverse-phase high-performance liquid chromatography and the N-terminal amino acid sequence of the purified protein was found to be identical to an internal portion of the deduced amino acid sequence of eas. Comparison of the sequences indicates a 29-amino-acid leader which is cleaved to generate the mature protein.
将粗糙脉孢菌分生孢子小梗制剂中的蛋白质溶解于三氟乙酸中。对溶解后的小梗进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,结果显示一条主要的蛋白质条带,其分子量约为7 kDa。携带eas突变的粗糙脉孢菌培养物制剂中不存在这种蛋白质。通过反相高效液相色谱法对该蛋白质进行纯化,发现纯化后蛋白质的N端氨基酸序列与eas推导氨基酸序列的内部部分相同。序列比较表明存在一个29个氨基酸的前导序列,该序列被切割后产生成熟蛋白质。