杨树SABP2基因的克隆与分析

发布时间：2018-05-18 07:57

本文选题：K杨 + 水杨酸结合蛋白(SABP)基因　；参考：《西北农林科技大学学报(自然科学版)》2017年03期

【摘要】：【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822bp,开放阅读框789bp,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。
[Abstract]:[objective] to clone and predict the function of 84K poplar salicylic acid binding protein (2(Salicylic acid-binding protein _ 2) SABP2. [methods] Total RNAs of leaves and stems of 84K poplar were extracted from 84 K poplar plantlets grown to 5 leaves. According to the complete CDs sequence of SABP2 gene of Populus tomentosa (GenBank sequence number: XM002310718.2), the primers of SABP2 gene of 84K poplar were designed, the whole length of SABP2 gene of 84K poplar was amplified by RT-PCR technique, then ligated to pGM-T clone vector, and transformed into E. coli TOP10 receptive cells. The SABP2 gene of 84 K poplar was cloned and its full length sequence was obtained. Bioinformatics analysis of the SABP2 gene and its encoded protein of 84K poplar was carried out by various online software. [results] the cDNA sequence of the 84K poplar gene was 822 BP, and the open reading frame 789 BP encoded 263 amino acids. Bioinformatics analysis showed that SABP2 and SABP2(GenBank had the highest homology. The SABP2 gene of Poplar 9884K had 11 protein binding sites in the microbody and belonged to the esterase of 伪 / 尾 folding hydrolase family. [conclusion] the SABP2 gene of poplar 84K was cloned successfully, and its function was consistent with the previous studies on the partial function of SABP2 in herbaceous plants.
【作者单位】：中国林业科学研究院森林生态环境与保护研究所国家林业局森林保护学重点实验室;南京林业大学南方现代林业协同创新中心;河南师范大学生命科学学院;中国林业科学研究院林业新技术研究所;
【基金】：中央级公益性科研院所基本科研业务费专项资金项目(CAFYBB2016SY037)
【分类号】：S792.11

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