杨树SABP2基因的克隆与分析

摘要

[Objective] This study cloned salicylic acid-binding protein 2 (SABP2) from 84K poplar and predicted its functions.[Method] The total RNA was isolated from foliage and stem of culture seedlings of 84K poplar with 5 leaves.The primers were designed according to the full CDs sequence of Populus trichocarpa SABP2 gene (Accession number:XM_002310718.2).The SABP2 full length of 84K poplar was amplified using RT-PCR.The product of PCR was ligated to pGM-T cloning vector and transformed into TOP10 competent cells chemically.Then,clone was obtained and sequenced.Bioinformatics analysis was also conducted by various online software.[Result] The full length cDNA sequence was 822 bp.The ORF was 789 bp,coding 263 amino acids.Bioinformatics analysis showed that the gene had 11 protein binding sites,and was homologous to populus tomentosa (JQ086570.1) with similarity of 98％.It was also a hydrophilic lipase belonging to the fold hydrolase super family.[Conclusion] The SABP2 gene of 84K poplar was cloned successfully.The predicted functions were consistent with previous research on herbs.%[目的]克隆84K杨水杨酸结合蛋白2(Salicylic acid-binding protein 2,SABP2)基因并预测其功能.[方法]以生长至5片小叶的84K杨组培苗为材料,提取其叶和茎的总RNA.根据毛果杨SABP2基因(GenBank序列号:XM_002310718.2)的完整CDs序列,设计84K杨SABP2基因的引物,采用RT-PCR技术扩增84K杨SABP2基因全长,然后连接到pGM-T克隆载体,转化大肠杆菌TOP10感受态细胞,对84K杨SABP2基因进行克隆,获得该基因的全长序列.通过各种在线软件对84K杨SABP2基因及其编码的蛋白质进行生物信息学分析.[结果]84K杨基因的cDNA序列全长822 bp,开放阅读框789 bp,编码263个氨基酸.生物信息学分析表明,84K杨SABP2与毛白杨SABP2(GenBank序列号:JQ086570.1)的同源性最高,达98％;84K杨SABP2基因位于微体中;SABP2蛋白有11个蛋白质结合位点,属于α/β折叠水解酶家庭成员中的酯酶,为亲水性蛋白.[结论]成功克隆了84K杨的SABP2基因,其功能与前人对草本植物中SABP2部分功能的研究结果一致.