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| 野桑蚕铜锌超氧化物歧化酶基因的克隆分析与原核表达 | |
| 引用本文： | 丛海峰,徐世清,司马杨虎,张升祥,王更先,董航,刘莹,柳学广.野桑蚕铜锌超氧化物歧化酶基因的克隆分析与原核表达[J].蚕业科学,2007,33(2):234-240. |
| 作者姓名： | 丛海峰 徐世清 司马杨虎 张升祥 王更先 董航 刘莹 柳学广 |
| 作者单位： | 1. 苏州大学生命科学学院,苏州,215123 2. 山东农业大学林学院,泰安,271018 3. 溧阳市蚕桑技术指导站,常州,213300 |
| 基金项目： | 国家自然科学基金项目(编号30371086)，国家重点基础研究发展计划“973”项目(编号2005CB121005) |
| 摘 要： | 超氧化物歧化酶是野桑蚕保护酶体系的重要酶类。利用RT-PCR方法克隆出野桑蚕铜锌超氧化物歧化酶(Cu/Zn-SOD)cDNA(EMB I登陆号:AM410997),其开放阅读框ORF长465 bp,编码154个氨基酸。同源性及系统进化分析表明,推导出的氨基酸序列与3种果蝇的同源性平均为69.3%,与线虫为57%,各物种中与Cu/Zn结合的残基高度保守。利用ExPASy的ScanProsite以及PSort和TMpred对此编码的蛋白质结构和功能域分析,预测出其具有2段Cu/Zn-SOD特异序列,且该蛋白不存在信号肽以及跨膜区。将Cu/Zn-SOD cDNA克隆到pET-28 a(+)表达载体,测序鉴定后以IPTG诱导表达,SDS-PAGE电泳鉴定其表达的融合蛋白质分子量为19.4 kD。 |
| 关 键 词： | 野桑蚕 铜锌超氧化物歧化酶 序列分析 原核表达 |
| 文章编号： | 0257-4799（2007）02-0234-07 |
| 修稿时间： | 2006-12-14 |
Cloning, Analysis and Expression of Copper/Zinc Superoxide Dismutase From Wild Silkworm, Bombyx mandarina |
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| CONG Hai-Feng,XU Shi-Qing,SIMA Yang-Hu,ZHANG Sheng-Xiang,WANG Geng-Xian,DONG Hang,LIU Ying,LIU Xue-Guang.Cloning, Analysis and Expression of Copper/Zinc Superoxide Dismutase From Wild Silkworm, Bombyx mandarina[J].Acta Sericologica Sinica,2007,33(2):234-240. | |
| Authors: | CONG Hai-Feng XU Shi-Qing SIMA Yang-Hu ZHANG Sheng-Xiang WANG Geng-Xian DONG Hang LIU Ying LIU Xue-Guang |
| Institution: | 1School of Life Science, Soochow University, Suzhou Jiangsu 215123, China; 2 College of Forestry, Shandong Agricultural University, Taian Shandong 271018, China ; 3 Sericultural Station of Liyang City, Changzhou Jiangsu 213300, China |
| Abstract: | Superoxide dismutase(SOD) is an important antioxygenic enzyme in Bombyx mandarina.This study focused on Copper/Zinc superoxide dismutase(Cu/Zn-SOD) from Bombyx mandarina.cDNA encoding Cu/Zn-SOD was firstly amplified by RT-PCR(EMBL accession number:AM410997).The sequence of cDNA indicated that the open reading frame comprises 465 base pairs encoding 154 amino acid residues.The deduced amino acid sequence of Cu/Zn-SOD showed 69.3% identity with those of three species of Drosophila on average,57% identity with that of Caenorhabditis elegans,respectively.The structure and functional domain of Cu/Zn-SOD was analyzed according to the software of ScanProsite,PSort and TMpred on line.The results showed that the protein contains two conservative motifs of Cu/Zn-SOD,and has no signal peptide or transmembrane region.The gene was cloned into expressing vector pET-28a( ) and expressed.SDS-PAGE analysis showed that the target protein was expressed at a high level and the putative molecular weight of recombinant protein was 19.4 kD. |
| Keywords: | Bombyx mandarina Copper/Zinc su-peroxide dismutase Sequence Analysis Prokary-otic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |