黑曲霉丝氨酸羧肽酶编码基因pepF的克隆、特性分析及表达

Cloning, characterization and expression of pepF, a gene encoding a serine carboxypeptidase from Aspergillus niger.

作者信息

van den Hombergh J P, Jarai G, Buxton F P, Visser J

机构信息

Section Molecular Genetics of Industrial Microorganisms, Wageningen Agricultural University, The Netherlands.

出版信息

Gene. 1994 Dec 30;151(1-2):73-9. doi: 10.1016/0378-1119(94)90634-3.

DOI:10.1016/0378-1119(94)90634-3

PMID:7828908

Abstract

We have cloned a gene (pepF) encoding a serine carboxypeptidase, proteinase F (PEPF), from Aspergillus niger. The sequences were identified in a phage lambda genomic DNA library using a synthetic probe based on the N-terminal sequence of PEPF. Nucleotide sequence data from pepF genomic and cDNA clones reveals that it is composed of four exons of 199, 283, 227 and 881 bp, interrupted by three introns of 53, 69 and 59 bp. The sequence of pepF codes for a polypeptide of 530 amino acids (aa), of which the first 52 aa are not present in the mature PEPF. This region may represent a prepro sequence that is removed by proteolytic cleavage as a monobasic cleavage site (Lys52). Northern blot analysis of total cellular RNA extracted from A. niger cells indicates that pepF is transcribed as a single 1.8-kb mRNA, which is regulated by nitrogen and carbon repression, specific induction and the pH of the culture medium.

摘要

我们从黑曲霉中克隆了一个编码丝氨酸羧肽酶即蛋白酶F（PEPF）的基因（pepF）。利用基于PEPF N端序列的合成探针，在λ噬菌体基因组DNA文库中鉴定出了这些序列。来自pepF基因组和cDNA克隆的核苷酸序列数据显示，它由199、283、227和881 bp的四个外显子组成，被53、69和59 bp的三个内含子打断。pepF的序列编码一个530个氨基酸（aa）的多肽，其中前52个氨基酸在成熟的PEPF中不存在。该区域可能代表一个前原序列，通过蛋白水解切割作为一个单碱性切割位点（Lys52）被去除。对从黑曲霉细胞中提取的总细胞RNA进行Northern印迹分析表明，pepF转录为一条1.8 kb的单链mRNA，其受氮和碳源阻遏、特异性诱导以及培养基pH值的调控。