Molecular cloning, characterization, and expression studies of a novel chitinase gene (ech30) from the mycoparasite Trichoderma atroviride strain P1.
We describe the cloning and characterization of a single copy gene from Trichoderma atroviride P1 encoding a novel 30 kDa chitinase, Ech30. Ech30 is a family 18 chitinase showing low sequence similarity to other Trichoderma chitinases. Real-time quantitative RT-PCR studies revealed that expression of the ech30 gene was induced by the presence of Botrytis cinerea in plate confrontation assays, but hardly by chitin in liquid cultures. Studies of Ech30 purified from an Escherichia coli strain overexpressing the ech30 gene devoid of the leader sequence and a predicted intron, showed that the gene encodes an active chitinase, which, as expected for family 18 chitinases, is inhibited by allosamidin.
Klemsdal
Sonja S
SS
Department of Plant Pathology, Plant Protection Centre, The Norwegian Crop Research Institute, Hoegskoleveien, Norway. sonja.klemsdal@planteforsk.no
Clarke
Jihong Liu
JL
Hoell
Ingunn A
IA
Eijsink
Vincent G H
VG
Brurberg
May-Bente
MB
D020121:5' Untranslated Regions / Q000235:genetics; D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D020171:Botrytis / Q000254:growth & development; D002686:Chitin / Q000494:pharmacology; D002688:Chitinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics; D014312:Trisaccharides / Q000494:pharmacology
D020121:5' Untranslated Regions; D004271:DNA, Fungal; D004791:Enzyme Inhibitors; D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014312:Trisaccharides; C054851:allosamidin; D002686:Chitin; D002688:Chitinases; D000117:Acetylglucosamine