棘孢木霉ACCC30536葡聚糖酶Glu41基因克隆及分析

Cloning and Sequence Analysis of Glucanase Gene Glu41 from Trichoderma asperellum ACCC30536

克隆获得生物防治菌棘孢木霉(T.asperellum)ACCC30536菌株葡聚糖酶基因Glu41的c DNA序列和DNA序列,cDNA和DNA的Gen Bank接受号分别为KJ541741和KJ541742,编码区长度为1134bp,编码378个氨基酸。克隆获得的启动子序列长度1500bp,含有6个逆境响应元件。Blast P相似性分析表明该基因与Glyco-hydrolase-16家族中棘孢木霉CBS433.97基因同源性最高相似度是60%,Signal P信号肽分析显示N端第25个和第26个氨基酸中间存在一个信号肽剪切位点(AFA||AP)。Pfam蛋白家族分析显示它属于Glyco-hydrolase-16家族的葡聚糖酶基因。将棘孢木霉ACCC30536菌株葡聚糖酶基因Glu41基因对Glyco-hydrolase-16家族中木霉属的CBS433.97基因组进行同源性搜索,在基因组上获得4个同源序列。4个同源基因表达中仅有Glu41在杨树根茎叶粉的诱导条件下表达。表明棘孢木霉葡聚糖酶基因Glu41与生物防治相关。

Glucanase gene Glu41 was cloned from the Trichoderma asperellum ACCC30536. The cDNA sequence of Glu41 is 1134 bp in length, encoding 378 amino acids. GenBank accession numbers of cDNA and DNA were KJ541741 and KJ541742, respectively. The promoter sequence was 1500bp in length and contained six stress response elements. BlastP search indicated that glucanase shared the highest similarity 60% with Glyco-hydrolase-16 superfamily of T. atroviride IMI 206040. At the same time, signalP prediction showed that the Glu41 amino acid sequence was cleaved by signal peptidase between positions 25 and 26 （AFAllAP）. Pfam protein family prediction showed that Glu41 protein belonged to Glyeo-hydrolase-16 superfamily. To obtain similarity gene sequences of glucanase Glu41 from the genome databases of T. asperellum CBS433.97, the amino acids sequences of Glu41 from T. asperellum ACCC30536 was used as query, resulting that 4 glucanase sequences with high similarity were obtained. Only Glu41 of those similarity genes expressed in the condition of inducing mixed powder of poplar root, stem and leaf. Above all, the study proved the glucanase gene Glu41 of T. asperellum was associated with plant disease biological control.