Ramón D, Carramolino L, Patiño C, Sánchez F, Peñalva M A
Departamento de Genética Molecular, Antibióticos S.A., Madrid, Spain.
Gene. 1987;57(2-3):171-81. doi: 10.1016/0378-1119(87)90120-x.
Genomic clones containing an Aspergillus nidulans isopenicillin N synthetase (IPNS) gene have been identified by heterologous hybridization with a Cephalosporium acremonium DNA probe. The open reading frame encodes a 331 amino acid polypeptide with extensive homology with the genes of other beta-lactam-producing fungi. The gene product has been overexpressed in Escherichia coli and shown to have activity of IPNS. This represents the first evidence at the molecular level that the biosynthesis of penicillins in A. nidulans occurs by the same pathway as in other beta-lactam-producing microorganisms. Comparison of available nucleotide sequences from IPNS genes suggests a horizontal transmission of the gene between the prokaryotic beta-lactam producers of the genus Streptomyces and the filamentous fungi.
通过与顶头孢霉DNA探针进行异源杂交,已鉴定出含有构巢曲霉异青霉素N合成酶(IPNS)基因的基因组克隆。该开放阅读框编码一个331个氨基酸的多肽,与其他产生β-内酰胺的真菌的基因具有广泛的同源性。该基因产物已在大肠杆菌中过表达,并显示具有IPNS活性。这代表了在分子水平上的首个证据,即构巢曲霉中青霉素的生物合成途径与其他产生β-内酰胺的微生物相同。来自IPNS基因的现有核苷酸序列比较表明,该基因在链霉菌属的原核β-内酰胺产生菌和丝状真菌之间存在水平转移。