枣树抗坏血酸过氧化物酶基因ZjAPX生物信息学分析及植物表达载体的构建  

The Biological Information Analysis and Construction of Plant Expression Vector of Zizyphus jujuba Ascorbate Peroxidase(APX)

作　　者：郝子琪[1,2] 许洪仙[3] 孟玉平[2] 曹冬梅[4] 曹秋芬[1,2] 

机构地区：[1]山西大学生物工程学院,山西太原030006 [2]山西省农业科学院生物技术研究中心,山西太原030031 [3]山西省农业科学院,山西太原030006 [4]山西省农业科学院园艺研究所,山西太原030031

出　　处：《山西农业科学》2011年第5期400-403,共4页Journal of Shanxi Agricultural Sciences

基　　金：山西省自然科学基金项目(2010011038-2);山西省财政支农专项

摘　　要：利用生物信息学软件对已获得的枣树抗坏血酸过氧化物酶基因cDNA序列ZjAPX进行了同源性及功能位点等多项参数分析。结果表明,此序列为全长753 bp的开放读码框(ORF),编码251氨基酸,分子量为62.55 kDa,理论等电点为5.10,为一个膜外蛋白;序列保守性分析表明,该序列具有典型的铁氧化还原蛋白结合区域,属于一个典型的植物亚铁血红素过氧化物酶家族蛋白;氨基酸序列相似性分析表明,该蛋白与已知的其他植物抗坏血酸过氧化物酶APX具有极高的同源性,与可可树APX和陆地棉APX同源性均为93%,命名为ZjAPX(DDBJ/EMBL/GenBank注册号:AB608053)。用Swiss Model程序(http://au.expasy.org/tools/)推测了ZjAPX基因编码蛋白的三维结构。以ZjAPX的cDNA序列为模板,PCR扩增后,经限制性内切酶消化,与PEZR(K)-LNY载体进行重组连接,测序结果显示,其植物表达载体构建成功。此结果为研究枣树抗坏血酸过氧化物酶基因在植物体内的生物学功能以及可能的活性调节方式奠定了基础。With obtained bioinformatics software, the homology and functional sites and many other parameters of the ascorbate peroxidase＇s eDNA sequences ZjA PX were analyzed. The results showed that this sequence was a protein of outside membrane and had an open reading frame （ORF） whose full-length was 753 bp, encoding 251 amino acids, its molecular weight 62.55 kDa; and theoretical isoelectric point 5.10. Conservation analysis of ZjAPX showed that it belonged to a plant heme peroxidase family with a typical zone of binding ferredoxin; similarity analysis showed that the protein had highly homologous with other known ascorbate peroxidase; the comparison between Theobroma cacao L., Gossypium hirsutum and ZjAPX was 93%. Therefore, it-was named ZjAPX （DDBJ/EMBL/GenBank：AB608053）. Tertiary structure of ZjA PX was speculated by Swiss Model program （http：//au.expasy.org/tools/）. Using the cloning vector pSPORT1 canoeing Zizyphus jujuba ascorbate peroxidase gene＇s cDNA sequence as a template to amplify the objective gene, then the PCR products were digested by BamH I and Sol I and connected to recombinant with plant expression plasmid PEZR （K）-LNY, and the sequencing results showed that the plant expression vector was successfully constructed. This data laid solid foundations for further research on the biological function of plant tissue and the possible ways of active regulation.

关 键 词：枣树 抗坏血酸过氧化物酶 生物信息学分析 植物表达载体 

分 类 号：S665.1[农业科学—果树学]