Cloning，Bioinformatics and Expression Analysis of UGDH1 Gene in Glycyrrhiza uralensis
10.13422/j.cnki.syfjx.20210246
- VernacularTitle:甘草UGDH1基因的克隆、生物信息学及表达分析
- Author:
Yi-fan SUN
1
;
Guang-xi REN
1
;
Ping LI
1
;
Dan JIANG
1
;
Chun-sheng LIU
1
Author Information
1. School of Chinese Materia Medica，Beijing University of Chinese Medicine，Beijing 102488，China
- Publication Type:Research Article
- Keywords:
Glycyrrhiza uralensis;
UGDH1 gene;
cloning;
bioinformatics;
polymerase chain reaction（PCR）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(12):133-140
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone uridine diphosphate （UDP）-glucose dehydrogenase （UGDH） gene of Glycyrrhiza uralensis and analyze its bioinformatics and expression. Method:Total RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of G. uralensis， the complementary deoxyribonucleic acid （cDNA） sequence of GuUGDH1 gene （Gu was short for G. uralensis） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）. Result:The open reading frame（ORF）of GuUGDH1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the GuUGDH1 gene was closely related to Glycine max and Spatholobus suberectus. The results of Real-time PCR showed that the expression of GuUGDH1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of G. uralensis， and the expression level in the roots was significantly higher than that in the stems and leaves. Conclusion:In this study， the UGDH1 gene of G. uralensis was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.
