朝鲜碱茅甜菜碱醛脱氢酶cDNA 5′末端序列的克隆及植物表达载体的构建  

Cloning of 5' cDNA Ends of Betaine Aldehyde Dehydrogenase from Puccinellia chinampoensis and Construction of Plant Expression Vector

作　　者：王磊[1] 钟鸣[1] 郭志富[1] 张丽[1] 张佳[1] 赵莉[1] 李浩戈[1] 

机构地区：[1]沈阳农业大学辽宁省农业生物技术重点实验室,辽宁沈阳110161

出　　处：《华北农学报》2010年第2期30-34,共5页Acta Agriculturae Boreali-Sinica

基　　金：辽宁省农业生物技术重点实验室专项基金

摘　　要：采用改良的RT-PCR及5′RACE法,成功地克隆了朝鲜碱茅甜菜碱醛脱氢酶(BADH)基因cDNA 5′端序列,与已知序列拼接,获得了BADHcDNA全长1781bp,包含一个完整的开放阅读框(ORF),编码506个氨基酸。其含有醛脱氢酶高度保守序列VT/SLELGGKSP,及其后29位与酶功能有关的Cys。多序列比对和系统进化分析表明,朝鲜碱茅BADH与大麦BBD1同源性最高,并构建了PBI121-BADH植物表达载体。The sequence of 5＇ eDNA ends of Betaine-aldehyde Dehydrogenase（BADH） was cloned from Puccinel- lia chinarnpoensis by improved RT-PCR and 5＇ RACE methods. The full length of eDNA was 1 781 bp by overlapping sequence and contained an intact open reading frame（ORF）encoding a putative protein of 506 amino acids. The de- duced amino acid sequence contained the conserved motif of aldehyde dehydrogenase which was VT/SLELGGKSP and after it there was the Cys at the 29th site,associated with aldehyde dehydrogenase function. Multiple sequence align- ment and phylogenetic analysis showed that the polypeptide shared high homology with Hordeum brevisubulatum BBD1. Then its plant expression vector named PBI121-BADH was constructed.

关 键 词：朝鲜碱茅 甜菜碱醛脱氢酶 5’RACE 高GC 表达载体 

分 类 号：Q785[生物学—分子生物学]