中间偃麦草TiAP1基因的克隆及生物信息学分析

Cloning and Bioinformatics of TiAP1 Gene from Thinopyrum intermedium

本研究从烟农15和中间偃麦草杂交所得的小偃麦异附加系SN6306中克隆得到TiAP1基因的全长序列,该序列全长1 272 bp,预测编码423个氨基酸。通过Blast P分析,发现其属于天冬氨酸蛋白酶家族。生物信息学分析发现,TiAP1蛋白含有信号肽且位于第16和17个氨基酸之间,同时含有两个木聚糖酶抑制剂结构域,分别位于该蛋白的N端和C端。此外,通过TMHMM预测,发现该蛋白无跨膜螺旋结构,因此是胞外蛋白。为了检测基因的亲缘关系,构建系统进化树,发现TiAP1基因与乌拉尔图小麦以及粗山羊草相应基因有较近的亲缘关系。上述结果为研究TiAP1基因的功能奠定了基础。

The wheat-Thinopyrum alien addition line SN6306 was originated from the intergeneric hybridization between YN15 and Thinopyrum intermedium（ Host） Nevski. Through PCR amplification,the full length c DNA of TiAP1 gene was obtained. Its open reading frame（ ORF） was 1 272 bp and encoded a peptide of 423 amino acids. The Blast P analysis results showed that it belonged to the aspartic protease family. The bioinformatics analysis results showed that TiAP1 protein contained a signal peptide,whose cleavage site was located between the 16 th and 17 th amino acid,and two xylanase inhibitor domain structures,which located in the N and C end respectively. The TMHMM predicting results showed that the protein had no transmembrane helix structure. So the protein was not secreted protein. The phylogenetic tree showed that TiAP1 gene was similar to the corresponding gene in Triticum urartu Thum. ex Gandil and Aegilops tauschii. These results would provide the reference for the research of TiAP1 gene functions.