Achlya bisexualis丙酮酸激酶基因的筛选、克隆及序列分析

Screening, Cloning and Sequence Ananlysis of the Gene for Pyruvate Kinase from Achlya bisexualis cDNA Library

从真菌藻状菌纲Achlyabisexualis的cDNA文库中克隆了糖酵解途径的最后一种酶———丙酮酸激酶的基因 ,获得这个基因的完整阅读框架 ,并且对这个基因的全序列进行了测定。使用在线计算机PSORTⅡ程序分析推导的氨基酸序列 ,发现此基因编码的丙酮酸激酶可能位于Achlyabisexualis的细胞质中。结果表明 ,在低等真核细胞的糖酵解途径的整个过程可能发生在细胞质中 ,与Liaud等人[1] 在硅藻细胞中的发现有所不同。

The gene for pyruvate kinase, which is involved in glyco ly sis and catalyze the biochemical reaction from phosphoenolpyruvate to pyruvate, was screened, cloned and sequenced from nonphotosynthetic Oomycetes, Achlya bi sexualis cDNA library. The size of the gene is about 1.6kb. The amino acid seq uence of the gene was deduced using the GCG program. On basis of analysis by the on-line PSORT Ⅱ program the deduced amino acid sequence showed that the pyruv ate kinase expressed by the gene was located in the cytosol. This result indicat ed that it is possible that the process of glycolysis for the lower form of euka ryotic cell such as Achlya bisexuali may occur in cytosol, which is differen t from Liaud's report .