观赏凤梨烯醇酶基因的克隆及分析

烯醇酶催化着糖酵解途径中的唯一一步脱水反应,与植物的抗逆性有着密切的联系.基于前期从擎天凤梨全长cDNA文库中获得的烯醇酶基因的EST单克隆,进行引物步移测序,获得其全长cDNA序列.序列长1 703bp,编码445个氨基酸残基,命名为GoEnolase1(GenBank登录号JN896863),蛋白质理论分子质量为47.9kD,等电点为5.7,包含1个TIM磷酸结合超家族保守区,二级结构主要由α螺旋(44.49%)、随机卷曲(33.71%)、延伸链(13.71%)和β转角(8.09%)组成.系统进化树分析表明,GoEnolase1蛋白与水稻、玉米、油棕等的烯醇酶蛋白聚为一类,它们的亲缘关系最近.

Cloning and Sequence Analysis of Enolase Gene from Ornamental Bromeliads

Enolase catalyze only one-step dehydration reaction in glycolytic pathway,and relate closely to plant stress resistance.Based on EST monoclones of enolase gene obtained from bromeliads full length cDNA library of ornamental bromeliads,its full length cDNA sequence was obtained by Primer Walking sequencing.The gene,named for GoEnolase1(GenBank accession No.JN896863),consisted of 1 703 bp cDNA sequence,1 335 bp ORF(open reading frame) which encoded a protein with 445 amino acids residues,and a putative protein which was 47.9 kD at estimated molecular weight,5.7 at isoelectric point and had a ’TIM-phosphate-binding superfamily’ conservative domain.The secondary structure of the protein was composed of alpha helix(44.49%),random coil(33.71%),extend strand(13.71%) and beta turn(8.09%) by SPOMA analysis.Furthermore,its tertiary structure was also build based on A chain of 2PSN.Through phylogenetic tree analysis,GoEnolase1 was gather to a same group with enolase protein in Oryza sativa,Zea mays and Elaeis guineensis.The research was beneficial to breeding for stress resistance in ornamental bromeliads.