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Image Search Results
Journal: Plant Physiology
Article Title: Decreased Protein Abundance of Lycopene β -Cyclase Contributes to Red Flesh in Domesticated Watermelon
doi: 10.1104/pp.19.01409
Figure Lengend Snippet: Catalytic function and localization of three ClLCYB proteins. A, Three ClLCYB haplotypes found in watermelons with different flesh colors: ClLCYBred with Val (V) at residue 226 and Lys (K) at residue 435 in all the red-fleshed lines; ClLCYByellow with Phe (F) at residue 226 and Asn (N) at residue 435 in the cultivated lines that did not accumulate lycopene, such as cv CS and cv JLM; and ClLCYBwhite with Phe (F) at residue 226 and Lys (K) at residue 435 in wild and semiwild white-fleshed watermelons. B, Subcellular localization of three ClLCYBs. ClLCYB-mCherry (ClLCYBwhite, ClLCYByellow, and ClLCYBred) and mCherry vector (vector control) were transiently produced in watermelon fruit protoplasts incubated in the dark for 12 h. The protoplasts were observed under chromoplast autofluorescence (column 1), under bright field (column 2), under mCherry fluorescence (column 3), and in merged images (column 4) with a confocal microscope. Bars = 20 µm. C, HPLC analysis of the color complementation of a pAC-LYC bacterial line with three ClLCYB constructs. The pAC-LYC plasmid (Cunningham and Gantt, 2007) enables lycopene production and accumulation in E. coli. pET28a expression vectors (Promega) carrying the three ClLCYB constructs were transformed into pAC-LYC cells, with empty pET28a as a control. Carotenoid pigment composition was examined in cultures of E. coli containing the plasmid indicated above. HPLC detection was performed at 440 nm. D, Michaelis-Menten profiles for the activities of three ClLCYBs and the concentration of the substrate lycopene in 5 mm NADPH, pH 6.5, at 25°C.
Article Snippet: The pAC-LYC plasmid ( Cunningham and Gantt, 2007 ) enables lycopene production and accumulation in E. coli .
Techniques: Residue, Plasmid Preparation, Control, Produced, Incubation, Fluorescence, Microscopy, Construct, Expressing, Transformation Assay, Concentration Assay
Journal: Frontiers in Microbiology
Article Title: The Protocatechuate 3,4-Dioxygenase Solubility (PCDS) Tag Enhances the Expression and Solubility of Heterogenous Proteins in Escherichia coli
doi: 10.3389/fmicb.2021.779541
Figure Lengend Snippet: Determination of the expression of recombinant PcaHG in E . coli BL21 (DE3) by SDS-PAGE. Total cell extract and soluble cell extract from induced E . coli BL21 (DE3). Lane M, protein molecular mass standards (Unstained Protein Molecular Weight Marker, No. 26610, Thermo Fisher Scientific, Waltham, MA, United States). (A) lane 1, total BL21 (DE3)/pET-28a (+), lane 2, soluble BL21 (DE3)/pET-28a (+), lane 3, total BL21 (DE3)/pET-28a- pcaHG98e01 , lane 4, soluble BL21 (DE3)/pET-28a- pcaHG98e01 , lane 5, total BL21 (DE3)/pET-28a- pcaHG98e02 , lane 6, soluble BL21 (DE3)/pET-28a- pcaHG98e02 . (B) lane 1, total BL21 (DE3)/pET-28a (+), lane 2, soluble BL21 (DE3)/pET-28a (+), lane 3, total BL21 (DE3)/pET-28a - pcaHG98e03 , lane 4, soluble BL21 (DE3)/pET-28a- pcaHG98e03 , lane 5, total BL21 (DE3)/pET-28a- pcaHG98e04 , lane 6, soluble BL21 (DE3)/pET-28a- pcaHG98e04 . (C) lane 1, soluble BL21 (DE3)/pET-28a- pcaHG98e04 ; lane 2. Purification of His 6 -PcaHG98e04-PCDS protein from BL21 (DE3)/pET-28a- pcaHG98e04 .
Article Snippet:
Techniques: Expressing, Recombinant, SDS Page, Molecular Weight, Marker, Purification
Journal: Frontiers in Microbiology
Article Title: The Protocatechuate 3,4-Dioxygenase Solubility (PCDS) Tag Enhances the Expression and Solubility of Heterogenous Proteins in Escherichia coli
doi: 10.3389/fmicb.2021.779541
Figure Lengend Snippet: Determination of pcaHG98 -encoded PCA 3,4 dioxygenase activity. Spectrophotometric changes during oxidation of PCA by pcaHG98 -encoded PCA 3,4 dioxygenase. The assay of PCA catalyzed by cell extracts from E . coli BL21 (DE3) carrying pET-28a- pcaHG98e01 (A) or pET-28a- pcaHG98e04 (D) . There was no enzyme activity in the cell extracts from BL21 (DE3) carrying pET-28a- pcaHG98e02 (B) or pET-28a- pcaHG98e03 (C) .
Article Snippet:
Techniques: Activity Assay