巴西橡胶树HbEBP1生物学特性、原核表达及氨肽酶活性分析  

The Characterization, Prokaryotic Expression and MetAP Activity Analysis of Hevea brasiliensis HbEBP1

作　　者：程汉[1] 祝建顺[1] 安泽伟[1] 方家林[1] 胡彦师[1] 黄华孙[1] 

机构地区：[1]中国热带农业科学院橡胶研究所,海南儋州571737

出　　处：《热带作物学报》2016年第2期317-324,共8页Chinese Journal of Tropical Crops

基　　金：农业部热带作物种质资源保护项目--橡胶树种质资源保护(No.15RZZY-01)

摘　　要：从巴西橡胶树中筛选到了Hb EBP1基因,该基因序列长度为1 462 bp,其中5′端非编码区长度为141 bp,3′端非编码区长度为133 bp,其c DNA序列具有完整的阅读框,编码395个氨基酸,推导的Hb EBP1分子量为43.596 ku,等电点位为6.45。生物信息学分析结果表明巴西橡胶树Hb EBP1具有3个保守结构域:增值相关蛋白2G4(PA2G4-like)、氨肽酶M24(APP_Met AP)、甲硫氨酸氨肽酶(MAP)结构域。对原核表达的Hb EBP1蛋白进行氨肽酶活性测定,发现体外表达的Hb EBP1蛋白没有Map活性。这些结果表明,橡胶树Hb EBP1基因并不存在氨肽酶活性。An EBP1 gene was identified from rubber tree in this study. The full length of the c DNA sequence of Hb EBP1 gene was 1 462 bp in length, with a 141 bp 5′ UTR and a 133 bp 3′ UTR. An intact ORF was found in the Hb EBP1 c DNA sequence, which encoding a 395 AA poly-peptide. The deduced Hb EBP1 protein had a molecular weight of 43.596 ku, p I 6.45. Bioinformatics analysis revealed this protein had three conserved domains:a PA2G4-like domain, a APP-Met AP domain and a MAP domain. Hb EBP1 was expressed in E.coli, and the purified protein was used to test the MAP activity. The results showed that Hb EBP1 protein had no MAP activity in vitro. These results demonstrated that Hb EBP1 may not function as a MAP enzyme.

关 键 词：橡胶树 HbEBP1 氨肽酶 原核表达 

分 类 号：S794.1[农业科学—林木遗传育种] Q78[农业科学—林学]