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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 被引量:1

Cloning of 5' cDNA Ends of Betaine Aldehyde Dehydrogenase from Puccinellia chinampoensis and Construction of Plant Expression Vector
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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.
出处 《华北农学报》 CSCD 北大核心 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
关键词 朝鲜碱茅 甜菜碱醛脱氢酶 5’RACE 高GC 表达载体 Puccinellia chinampoensis BADH 5' RACE GC-rich Expression vector
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