Open AccessDetection of putative peptide synthetase genes in Trichoderma species: Application of this method to the cloning of a gene from T. harzianum CECT 2413
Author(s) -
Vizcaíno J.A.,
Sanz L.,
Cardoza R.E.,
Monte E.,
Gutiérrez S.
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.01.036
Subject(s) - adenylylation , biology , gene , trichoderma harzianum , pseudogene , open reading frame , microbiology and biotechnology , peptide , cloning (programming) , genetics , biochemistry , peptide sequence , biosynthesis , genome , ecology , biological pest control , computer science , programming language
Some of the secondary metabolites produced by Trichoderma , such as the peptaibols and other antibiotics, have a peptide structure and in their biosynthesis are involved proteins belonging to the Non‐Ribosomal Peptide Synthetase family. In the present work, a PCR‐mediated strategy was used to clone a region corresponding to an adenylation domain of a peptide synthetase (PS) gene from 10 different strains of Trichoderma . In addition, and using the fragment isolated by PCR from T. harzianum CECT 2413 as a probe, a fragment of 19.0 kb corresponding to a PS‐encoding gene named salps1 , including a 1.5 kb fragment of the promoter, was cloned and sequenced. The cloned region of salps1 contains four complete, and a fifth incomplete, modules, in which are found the adenylation, thiolation and condensation domains, but also an additional epimerization domain at the C‐terminal end of the first module. The analysis of the Salps1 protein sequence, taking into consideration published data, suggests that it is neither a peptaibol synthetase nor a protein involved in siderophore biosynthesis. The presence of two breaks in the open reading frame and the expression of this gene under nitrogen starvation conditions suggest that salps1 could be a pseudogene.