Cloning and Bioinformatics Analysis of Chitin Synthase Gene ThChsC from Trichoderma harzianum

【Objective】 The objective of this study is to clone and make bioinformatics analysis of the sequence of chitin synthase gene ThChsC from Trichoderma harzianum.【Method】 The full length of ThChsC was cloned by polymerase chain reaction(PCR) and genome walking methods,the gene sequence obtained and the putative amino acid sequence were analyzed by bioinformatics software,and the three-dimensional structure of ThChsC was predicted by homology modeling.【Result】 ThChsC(GenBank accession number HQ419000) coding sequence spans 2 835 bp and is comprised of 4 extrons inserted by 3 introns.The ThChsC contains an open reading frame(ORF) of 2 688 bp capable of encoding a polypeptide of 895 amino acids and shares 80% and 81% identity to Gibberella moniliformis chsB(ACY08039.1) and Colletotrichum graminicola chs2(AAL23718.1),respectively.Phylogenetic analysis indicates that ThChsC belongs to the type III chitin synthase subfamily.The conserved Chitin_synth_1,Chitin_synth_1N and Chitin_synth_2 domains and three transmembrane domains were found in the predicted ThChsC protein.A conserved DHD domain of glycosyl transferase was found in the three-dimensional structure of ThChsC predicted.【Conclusion】The chitin synthase gene ThChsC was cloned for the first time from Trichoderma harzianum Th-33.The further study of ThChsC will have an important significance in exploring the mechanism of chitin synthesis of Trichoderma harzianum.