棘孢木霉ACCC30536葡聚糖酶基因的克隆及表达

以棘孢木霉(Trichoderma asperellum)ACCC30536菌株为试材,采用聚合酶链式反应(PCR)技术克隆获得葡聚糖酶GH71基因的全长c DNA序列。结果表明:此基因的c DNA编码区序列全长1 296 bp,编码区可编码431个氨基酸。经Blast P相似性分析,其属于GH99~GH71超家族,与深绿木霉(T.atroviride)氨基酸序列XP_013941146的同源性最高,达到94%。7种不同诱导条件下,GH71基因的表达量变化明显。在2种病原菌细胞壁的诱导下,GH71基因的表达均呈先上升后下降又上升的趋势,在培养的12 h时达到最大值,分别为未诱导时的24.3倍和23.9倍。而在这2种病原菌发酵液的诱导下,GH71基因的表达均呈先下降后上升的趋势。在山新杨茎与叶的诱导下,GH71基因的表达量在24 h最大,分别为未诱导的25.4倍和25.2倍;在山新杨根的诱导下,GH71基因的表达量在48 h达到最大,是未诱导的28.1倍。

Cloning and Differential Expression of A Glucanase Gene from Trichoderma asperellum ACCC30536

We cloned and isolated the cDNA of glucanase (GH71) gene from Trichoderma asperellum ACCC30536.The cDNA sequence of GH71 was 1296 bp in length, encoding 431 amino acids.BlastP search indicated that GH71 belonged to GH99-GH71-like superfamily and it showed the highest similarity 94%with a protein of T.atroviride ( XP_013941146 ) . Moreover , GH71 was proven to be differentially transcribed under seven different treatments .GH71 transcripts were up-reg-ulated in the conditions of inducing both 1%Alternaria alternata cell wall and 1%Cytspora chrysosperma cell wall with peak transcription levels of 24.3 and 23.9 times at 12 h, respectively.While under the conditions of 10%A.alternata fer-mentation liquid and C.chrysosperma fermentation liquid , the transcripts of GH71 were down-regulated at first and then up-regulated.GH71 transcription was up-regulated by 1%leaf and stem powder of Ppo ulus dividiana×P.bolleana with peak transcription levels of 25.4 and 25.2 times at 24 h, respectively .The transcript of GH71 was also up-regulated by 1%root of Populus dividiana×P.bolleana, with peak transcription levels of 28.1 times at 48 h.