Vastag Mária, Kasza Zsolt, Acs Klára, Papp Tamás, Schwab Helmut, Vágvölgyi Csaba
Department of Microbiology, Faculty of Sciences, University of Szeged, P.O. Box 533, H-6701 Szeged, Hungary.
Antonie Van Leeuwenhoek. 2004 Aug;86(2):111-9. doi: 10.1023/B:ANTO.0000036118.41484.e7.
Rhizomucor miehei is important from a biotechnological aspect in consequence of its content of aspartic proteinase, which has high milk-clotting activity. A genomic library of R. miehei NRRL 5901 has been constructed in a phage (Lambda Fix II) vector. The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene was isolated from this library by hybridization of the recombinant phage clones with a gpd-specific gene probe generated by polymerase chain reaction. The complete nucleotide sequence encodes a putative polypeptide chain of 336 amino acids interrupted by 5 introns. The predicted amino acid sequence of this gene shows a high degree of sequence similarity to the glyceraldehyde-3-phosphate dehydrogenase proteins from yeast and filamentous fungi. The promoter region, containing a consensus TATA box, and 246-bp downstream from the putative stop codon were also determined. The possibility of using the gpd promoter in the construction of new transformation vectors is discussed.
米黑根毛霉因其天冬氨酸蛋白酶含量而在生物技术方面具有重要意义，该蛋白酶具有很高的凝乳活性。已使用噬菌体（λFix II）载体构建了米黑根霉NRRL 5901的基因组文库。通过重组噬菌体克隆与聚合酶链反应产生的gpd特异性基因探针杂交，从该文库中分离出甘油醛-3-磷酸脱氢酶（gpd）基因。完整的核苷酸序列编码一个由336个氨基酸组成的推定多肽链，该多肽链被5个内含子打断。该基因预测的氨基酸序列与酵母和丝状真菌的甘油醛-3-磷酸脱氢酶蛋白具有高度的序列相似性。还确定了含有共有TATA框的启动子区域以及推定终止密码子下游246 bp的区域。讨论了在构建新的转化载体中使用gpd启动子的可能性。
