毛竹PsbS基因的克隆及其原核表达特征研究  

Cloning of PsbS Gene from Moso Bamboo(Phyllostachys edulis) and Its Characterization of Prokaryotic Expression

作　　者：高志民[1] 于小清[1,2] 郑波[1] 李雪平[1] 胡陶[1] 

机构地区：[1]国际竹藤网络中心,国家林业局竹藤科学与技术重点开放实验室,北京100102 [2]美国普渡大学农学系,美国印第安纳州47907

出　　处：《热带亚热带植物学报》2011年第6期513-518,共6页Journal of Tropical and Subtropical Botany

基　　金：国家自然科学基金项目(30972328);林业公益性行业科研专项(200704001);国际竹藤网络中心基本科研业务费专项基金(1632010004)资助

摘　　要：采用RT-PCR技术从毛竹(Phyllostachys edulis)叶片中克隆到1个PsbS基因,命名为PePsbS(GenBank No.FJ600727),其编码区为810 bp,编码269个氨基酸。序列分析表明,PePsbS编码的蛋白与其它单子叶植物的PsbS蛋白有很高的相似性。蛋白结构分析表明,PePsbS基因编码蛋白包含导肽部分和成熟蛋白,其中成熟蛋白包含4个跨膜结构域。将PePsbS基因编码成熟蛋白的序列构建到原核表达载体pET23a中,并转入大肠杆菌,用IPTG进行诱导表达。结果表明,41℃下诱导4 h的表达效果最好,目的蛋白含量约占总蛋白的21.5%,分子量约为22.0 kD。这说明温度和诱导时间明显影响PePsbS基因的表达。PsbS protein plays an important role in non-photochemical quenching （NPQ） in plants. A PsbS gene was cloned from Moso bamboo （Phyllostachys edulis） leaves by RT-PCR method, which named PePsbS （GenBank No. FJ6（13727） with 810 bp in length, and encoded a polypeptide with 269 amino acids. Horrology analysis showed that the deduced PsbS was highly homologous to those from other tronocotyledon plants. The protein structure analysis indicated that PePsbS consists of transit peptide and mature protein including four transmembrane domains. The fraglrent ofPePsbS gene encoding mature protein was constructed in rmltiple cloning site ofpETZ3a and expressed in Eschedchia coli induced by IPTG. The expression of PePsbS was the best under 41℃ for 4 hours, the recombinant protein accounted for 21.5% of the total proteins with molecular weight about 22 kD. It indicated that there were significant effects of terrpemture and IPTG induced time on protein expression.

关 键 词：毛竹 PsbS基因 克隆 载体构建 原核表达 温度 

分 类 号：S795[农业科学—林木遗传育种]