1.Cloning, prokaryotic expression, and functional validation of flavonoid 3-O-glycosyltransferase gene (Rh3GT) from Rhododendron hybridum Hort.
Yicheng YAN ; Zehang WU ; Yuhang JIANG ; Gaoyuan HU ; Yujie YANG ; Xiaohong XIE ; Yueyan WU ; Yonghong JIA
Chinese Journal of Biotechnology 2025;41(2):881-895
Flavonoid 3-O-glucosyltransferase (3GT) is a key enzyme in the glucosidation of anthocyanins. To investigate the 3GT gene in rhododendron, we cloned an open reading frame (ORF) of 3GT gene (named Rh3GT) from Rhododendron hybridum Hort (Red cultivar) and then characterized this gene and the deduced protein in terms of the biochemical characteristics, expression level, and enzymatic function. The results showed that Rh3GT had a full length of 993 bp and encoded 330 amino acid residues. The deduced protein was hydrophilic, stable, weak acid, belonging to the glycosyltransferase family (GT-B type), with glutamine (Q) at position 44 in the PSPG box. The phylogenetic analysis showed that Rh3GT was most closely related to Vc3GT from Vaccinium corymbosum and Vm3GT from Vaccinium myrtillus. Rh3GT was expressed in the stems, leaves, and flowers and almost not expressed in the roots, with the highest expression level in petals during full blooming stage. Introduction of pCAMBIAL1302-Rh3GT into petals significantly up-regulated the expression level of Rh3GT and increased the total anthocyanin accumulation. Rh3GT was successfully expressed in Escherichia coli BL21 in the form of inclusion bodies with a size of about 36 kDa. The results of HPLC showed that the recombinant Rh3GT after denaturation, purification, and dilution could catalyze the synthesis of cyanidin and UDP-glucose to synthesize cyanidin 3-O-glucoside, indicating that the expressed protein had 3GT activity. This study provides basic data for further studying the molecular regulation mechanism of anthocyanin biosynthesis and theoretical support for molecular breeding of rhododendron.
Rhododendron/classification*
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Glucosyltransferases/metabolism*
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Cloning, Molecular
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Escherichia coli/metabolism*
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Recombinant Proteins/biosynthesis*
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Anthocyanins/biosynthesis*
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Phylogeny
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Plant Proteins/metabolism*
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Amino Acid Sequence
2.Early warning of high altitude pulmonary edema
Gaoyuan LI ; Zongbin LI ; Jinwu ZHANG ; Junjie JIANG ; Anzhong HU ; Anheng LIU
Military Medical Sciences 2015;(4):254-256
Objective By analyzing the clinical symptoms, heart rate ( HR) , arterial oxygen saturation ( SaO2 ) and the number of white blood cells (WBC), we aimed to explore the implication of the above-mentioned indexes for early warning of high altitude pulmonary edema ( HAPE) .Methods Based on the Lake Louise Self-assessmeat Scoring System ( LLSS) and the scoring of respiratory symptoms, 628 subjects were divided into three groups: group A ( the healthy;score<3),group B(acute mountain sickness, AMS; score>3 and excluding HAPE),and group C (HAPE).Moreover, we analyzed the incidence of some clinical symptoms, HR and SaO2 , as well as the WBC number of some subjects in the three groups.Results The incidence of respiratory symptoms and WBC number were significantly increased in group C compared with group B(P<0.05).In addition,unlike group A and B, the mean HR in group C rose gradually in the first 3 days and was markedly increased at the third day(F=6.37,P<0.05).The mean SaO2in group C was remarkably lower than in group A and B in the first 3 days(F=8.21,F=8.77,F=9.58,P<0.01).Conclusion Those who enter high altitude for the first time with notable respiratory symptoms, WBC increase, HR maladaptation and decrease in SaO2 (>30%) have high risk of HAPE.It is of special importance to detect HAPE earlier at high altitude.
3.A correlation study of Tei index and N-terminal pro-brain natriuretic peptide in patients with high altitude heart disease
Gaoyuan LI ; Zhengjian LIU ; Haijun CHEN ; Xuehong ZHANG ; Junjie JIANG ; Anzhong HU
Chinese Journal of Internal Medicine 2011;50(11):947-949
ObjectiveTo explore the diagnostic value of Tei index of right ventricle and serum level of NT-proBNP in patients with high-altitude heart disease (HAHD).MethodsRight ventricle Tei index and serum NT-proBN level were calculated and tested in 32 local healthy volunteers and 34 cases of HAHD patients hospitalized in our hospital in Golmud city (2808 meters above sea level) from 2008 to 2010,and a correlation study was conducted thereafter.ResultsThe pulmonary arterial systolic pressure and right ventricle Tei index,elevated significantly in HAHD patients compared with the control group [(86.61 vs 9.72)mm Hg(1 mm Hg=0.133 kPa) and(0.90 vs 0.33)respectively,P<0.05].Patients diagnosed as mild pulmonary hypertension without alteration in cardiac structure showed higher pulmonary arterial systolic pressure and the Tei index compared with the control group [( 57.1 vs 9.72 )mm Hg and (0.78 vs 0.33 ) respectively,P < 0.05 ].In addition,the level of serum NT-proBNP was significantly higher in HAHD group than that of control group [( 1246.8 ± 512.6) ng/L and (98.6 ± 21.7 ) ng/L respectively,P < O.05 ].ConclusionRight ventricle Tei index and serum NT-proBNP level are sensitive indicators for right ventricular function and thus of favorable clinical significance for the diagnosis of HAHD.

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