油桐ALDH22A1和ALDH2C4基因克隆及其序列分析

为给油桐ALDH基因的结构与功能研究提供理论依据,以葡萄桐的近成熟种子为材料,根据油桐转录组测序结果设计引物,采用RT-PCR技术克隆了油桐ALDH22A1基因和ALDH2C4基因的全长cDNA序列。ALDH22A1的cDNA序列全长1 782 bp,编码593个氨基酸,该基因编码蛋白质的相对分子质量为65.60 kDa,理论等电点为6.62,蛋白二级结构以α螺旋为主,具有1个明显的跨膜结构,是稳定的非分泌蛋白。ALDH2C4的cDNA序列全长1 506 bp,编码501个氨基酸,该基因编码蛋白质的相对分子质量为54.82 kDa,理论等电点为6.58,蛋白二级结构以α螺旋为主,是不具有跨膜结构的膜外蛋白。BLASTp分析结果表明,这2个基因所编码序列与麻疯树、蓖麻的乙醛脱氢酶一致性最高,高达80%以上,含有醛脱氢酶基因家族的保守结构域。

cDNA cloning and sequence analysis of ALDH22A1 and ALDH2C4 genes in Verniciafordii

In order to provide some theoretical basis for the studies on structure and function ofALDH gene in Vernicia fordii,using nearly ripe seeds in ‘Putaotong’ as materials,primers were designed according to the results of the transcriptome sequencing analysis of V..fordii,and full-length cDNA sequences of ALDH22A1 and ALDH2C4 genes were cloned by RT-PCR techniques.The cDNA sequence length ofALDH22A1 gene was 1 782 bp,encoding 593 amino acids.The relative molecular mass of ALDH22A1 protein was 65.60 kDa,and the theoretical isoelectric point was 6.62.The predicted second structure of the protein mainly consisted of alpha helix,and had one obvious transmembrane domain,and was a stable non-secretory protein.The cDNA sequence length ofALDH2C4 gene was 1 506 bp,encoding 501 amino acids.The relative molecular mass of ALDH2C4 protein was 54.82 kDa,and the theoretical isoelectric point was 6.58.The predicted second structure of the protein mainly consisted of alpha helix,and had no transmembrane domain.The results of Blastp analysis showed that,these sequences encoded by the two genes had the highest consistency with acetaldehyde dehydrogenase in Jatropha curcas and Ricinus communis,up to 80％,and contained the conserved domain of the aldehyde dehydrogenase gene family.