深绿木霉1-氨基环丙烷-1-羧酸脱氨酶基因的克隆及表达

摘要

The cDNA of ACC deaminase ( ACCD) gene was isolated from Trichoderma atroviride ACCC30153 by using PCR .The cDNA sequence of ACCD was 1083 bp in length , encoding 360 amino acids .BlastP search indicated that ACCD belonged to Try-synth-beta-Ⅱfamily and it shown the highest similarity 91%with a protein of T.reesei ( XP-006967764 ) .ACCD was proven to be differentially transcribed under five different treatments .ACCD transcripts were up-regulated under nutri-tional stress conditions with peak transcription levels of 22.7 and 22.4 times, at 8 h and 16 h, respectively.ACCD transcrip-tion was also up-regulated by 1%leaf and stem powder of Populus davidiana ×P.alba var.pyramidalis, with peak tran-scription levels of 22.6 and 27.3 times, at 24 h and 2 h, respectively.%以深绿木霉(Trichoderma atroviride)ACCC30153为试材,应用聚合酶链式反应(PCR)技术克隆获得1-氨基环丙烷-1-羧酸脱氨酶ACCD基因的cDNA全长.结果表明:此基因的cDNA编码区序列全长1083 bp,编码区可编码360个氨基酸.经BlastP相似性分析,其属于Try-synth-beta-Ⅱ家族,与里氏木霉(T.reesei)氨基酸序列XP-006967764的同源性最高,达到91%.在5种不同诱导条件下,ACCD基因的表达量均有明显变化.在C、N饥饿培养基中分别培养8 h与16 h时,ACCD基因表达量达到最大值,为未诱导时的22.7倍与22.4倍;在山新杨叶粉与茎粉的诱导下,ACCD的表达量分别在培养的24 h与2 h达到最大,为未诱导的22.6倍与27.3倍.