title: Isolation and functional analysis of spy1 responsible for pristinamycin yield in Streptomyces pristinaespiralis.
abstract: A gene related to high pristinamycin yield in Streptomyces pristinaespiralis was selected by amplified fragment length polymorphism (AFLP) and its functions were investigated by gene disruption. First, a 561 bp polymorphic sequence was acquired by AFLP from high-yield recombinants compared with the S. pristinaespiralis ancestor ATCC25486, indicating that this approach is an effective means of screening for valuable genes responsible for antibiotic yield. Then, a 2,127 bp open reading frame of a gene designated spy1 that overlaps with the above fragment was identified and its structure and biological functions were investigated. In silico analysis of spy1 encoding a deduced 708-amino-acid-long serine/threonine protein kinase showed that it only contains a catalytic domain in the N-terminal region, which is different from some known homologs. Gene inactivation of chromosomal spy1 indicated that it plays a pleiotropic regulatory function in pristinamycin production, with a positive correlation to pristinamycin I biosynthesis and a negative correlation to pristinamycin II biosynthesis.
authors.affiliation: School of Biological and Chemical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo 315100, China.
authors.forename: Qingchao
authors.initials: Q
authors.lastname: Jin
authors.forename: Huali
authors.initials: H
authors.lastname: Yin
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Hong
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Jin
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D005798:Genes, Bacterial; D016366:Open Reading Frames; D025762:Pristinamycin / Q000096:biosynthesis*; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D025762:Pristinamycin; D011494:Protein Kinases