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。
王磊
,
钟鸣
,
郭志富
,
张丽
,
张佳
,
赵莉
,
李浩戈
. 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[J]. 华北农学报, 2010
, 25(2)
: 30
-34
.
DOI: 10.7668/hbnxb.2010.02.006
The sequence of 5' cDNA ends of Betaine-aldehyde Dehydrogenase(BADH)was cloned from Puccinellia chinampoensis by improved RT-PCR and 5' RACE methods.The full length of cDNA was 1 781 bp by overlapping sequence and contained an intact open reading frame(ORF)encoding a putative protein of 506 amino acids.The deduced 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 alignment and phylogenetic analysis showed that the polypeptide shared high homology with Hordeum brevisubulatum BBD1.Then its plant expression vector named PBI121-BADH was constructed.
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