热带作物学报2017，Vol.38Issue(4)：673-681,9.DOI:10.3969/j.issn.1000-2561.2017.04.014
玳玳花瓣脂氢过氧化物裂解酶基因cDNA的克隆及原核表达
cDNA Cloning and Prokaryotic Expression of Hydroperoxide Lyase in Daidai (Citrus aurantium)
郭凤芝 1许颖妍 1熊青 1刘涛 1吕恃衡 1陈桂信 1佘文琴1
作者信息
- 1. 福建农林大学园艺产品贮藏保鲜研究所,福建福州 350002
- 折叠
摘要
Abstract
In this research,the full length cDNA of Hydroperoxide lyase (HPL) from Daidai (Citrus aurantium) petals was cloned by RT-PCR and RACE and tried to express this gene in E.coli.The cDNA sequence of Daidai HPL was 1 776 bp long.It contained an ORF encoding 499 amino acid residues (Mw=55.7 ku),a 5'untranslated region of 52 bp and a 3'untranslated region of 224 bp.Daidai HPL was a hydrophilic protcin.Compared with HPLs of other plants,this phylogenetic analysis suggested that Daidai HPL was a member of the 13-HPL family.Genomic fragment of Daidai HPL gene was also amplified by using PCR,and subsequent sequencing analysis showed that the Daidai HPL gene contained one intron of 2 374 bp.The Daidai HPL gene cDNA was inserted into the vector pET-15b.The expression construct was transformed into BL21 and was induced.Daidai HPL was expressed at high level in E.coli.SDS-PAGE analysis showed that Daidai HPL was 55 ku,which is approximately the same with the calculated molecular weight 55.7 ku.The research aimed to control the expression of HPL gene and regulate he production of scents in plants,and provide a new approach to the genetic improvement of flower scents.关键词
玳玳(Citrus aurantium)/脂氢过氧化物裂解酶/cDNA全长/原核表达Key words
Daidai (Citrus aurantium)/hydroperoxide lyase/full length cDNA/expression in E.coli.分类
生物科学引用本文复制引用
郭凤芝,许颖妍,熊青,刘涛,吕恃衡,陈桂信,佘文琴..玳玳花瓣脂氢过氧化物裂解酶基因cDNA的克隆及原核表达[J].热带作物学报,2017,38(4):673-681,9.
