Cloning and expression of SmDXS2 gene in Swertia mussotii.
10.19540/j.cnki.cjcmm.20181226.004
- Author:
Wen-Jing LI
1
;
Bei-Bei XIANG
2
;
Yan-Xiang SUN
1
;
Xiao-Qiang HOU
1
;
Mei-Ling HAN
1
;
Xiao-Xue LI
3
;
Yong WANG
3
;
Shuo GUO
1
Author Information
1. College of Life Science,Langfang Teachers University Langfang 065000,China.
2. School of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine Tianjin 300193,China.
3. College of Life Science,Nankai University Tianjin 300071,China.
- Publication Type:Journal Article
- Keywords:
1-deoxy-D-xylulose-5-phosphate synthase2;
Swertia mussotii;
bioinformations analysis;
gene cloning;
prokaryotic expression
- MeSH:
Amino Acid Sequence;
Cloning, Molecular;
DNA, Complementary;
genetics;
Genes, Plant;
Iridoids;
Phylogeny;
Plant Proteins;
genetics;
Swertia;
enzymology;
genetics;
Transcriptome;
Transferases;
genetics
- From:
China Journal of Chinese Materia Medica
2019;44(5):935-941
- CountryChina
- Language:Chinese
-
Abstract:
1-deoxy-D-xylulose-5-phosphate synthase2(DXS2) is the first key enzyme of the MEP pathway,which plays an important role in terpene biosynthesis of plants. According to the data of Swertia mussotii transcriptome, DXS2 gene(Gen Bank number MH535905) was cloned and named as Sm DXS2. The bioinformatics results showed that Sm DXS2 has no intron,with a 2 145 bp open reading frame encoding a polypeptide of 714 amino acids. They are belonging to 20 kinds of amino acids,and the most abundant amino acids include Ala,Gly and Trp. The predicted protein molecular weight was 76. 91 k Da and its theoretical isoelectric point(p I) was6. 5,which belonging to a hydrophilic protein. α-Helix and loop were the major motifs of predicted secondary structure of DXS2. The three function domains are TPP＿superfamily,Transket＿pyr＿ superfamily and Transketolase＿C superfamily,respectively. The Sm DXS2 protein shared high identity with other DXS2 proteins of plants. Phylogenetic analysis showed that Sm DXS2 protein is grouped with the gentian DXS2 protein. The recombinant protein of Sm DXS2 gene in Escherichia coli was approximately 92. 00 k Da(containing sumo-His tag protein 13 k Da),which was consistent with the anticipated size.This work will provide a foundation for further functional research of Sm DXS2 protein and increasing the product of iridoid compound by genetic engineering in S. mussotii.
