中国中药杂志

2019, v.44(15) 3253-3260

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

盐肤木查尔酮异构酶的克隆、表达及特性分析
Cloning,expression and characterization of chalcone isomerase from medicinal plant Chinese sumac (Rhus chinensis)

杨晓月;王景;李晨;任竹梅;马文丽;
YANG Xiao-yue;WANG Jing;LI Chen;REN Zhu-mei;MA Wen-li;School of Life Science,Shanxi University;

摘要(Abstract):

黄酮类化合物是药用植物盐肤木Rhus chinensis中主要的功能成分,具有多种药理活性,广泛用于疾病治疗和食品保健等领域。查尔酮异构酶(chalcone isomerase,CHI,EC 5. 5. 1. 6)是黄酮类化合物生物合成途径的关键酶。该研究通过RTPCR及rapid-amplification of c DNA Ends (RACE)技术克隆了盐肤木查尔酮异构酶(命名为Rc CHI)全长c DNA序列共1 058bp,包含738 bp的开放阅读框,编码245个氨基酸。序列分析表明,Rc CHI与其他植物CHI序列相似性在47. 1%~71. 6%,属于Ⅰ型CHI。通过实时荧光定量PCR检测发现盐肤木不同组织总黄酮含量与Rc CHI的mRNA表达呈正相关。进一步构建原核表达载体p GEX-6P-1-RcCHI并转化大肠杆菌BL21,成功获得了重组Rc CHI的高效可溶表达。该研究为利用基因工程菌株进行黄酮类化合物的生物合成奠定了基础。
Flavonoids are a group of secondary metabolites found in plants. They have many pharmacological functions and play an important role in Chinese sumac( Rhus chinensis),which is a well-known traditional Chinese medicinal plant. Chalcone isomerase( CHI,EC 5. 5. 1. 6) is one of the key enzymes in the flavonoids biosynthesis pathway. In this paper,the full-length c DNA sequence encoding the chalcone isomerase from R. chinensis( designated as Rc CHI) was cloned by RT-PCR and rapid-amplification of c DNA Ends( RACE). The Rc CHI c DNA sequence was 1 058 bp and the open reading frame( ORF) was 738 bp. The ORF predicted to encode a 245-amino acid polypeptide. Rc CHI gene contained an intron and two exons. The sequence alignments revealed Rc CHI shared47. 1%-71. 6% identity with the homologues in other plants. Real-time PCR analysis showed that the total flavonoid levels were positively correlated with tissue-specific expressions of Rc CHI mRNA in different tissues. The recombinant protein was successfully expressed in an Escherichia coli strain with the p GEX-6 P-1 vector. In this paper,the CHI gene was cloned and characterized in the family of Anacardiaceae and will help us to obtain better knowledge of the flavonoids biosynthesis of the flavonoid compounds in R. chinensis.

关键词(KeyWords): 盐肤木;查尔酮异构酶;分子克隆;表达;黄酮生物合成
Rhus chinensis;chalcone isomerase(CHI);cloning;expression;flavonoids biosynthesis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(31870366);; 山西省留学基金项目(2016-005)

作者(Author): 杨晓月;王景;李晨;任竹梅;马文丽;
YANG Xiao-yue;WANG Jing;LI Chen;REN Zhu-mei;MA Wen-li;School of Life Science,Shanxi University;

Email:

DOI:

参考文献(References):

扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享