Lactose, glucose, standard GOS, and gaLactose were supplied by Solarbio (Beijing, China). O-nitrophenyl-β-D-galactopyranoside (ONPG) and o-nitrophenol (ONP) were purchased from Sangon Biotech (Shanghai, China). Besides, the plasmid pET-28a (+) (Novagen, Madison, WI, USA) and the E. coli strain BL21 (DE3) (Tiangen Biotech, Beijing, China) were used for plasmid construction and gene expression host, respectively. The silica gel thin-layer chromatography (TLC) plates were purchased from Merck (Darmstadt, Germany). All reagents and chemicals were of analytical grade unless stated otherwise.
O nitrophenyl β d galactopyranoside
Manufactured by Sangon
Sourced in China
O-nitrophenyl-β-D-galactopyranoside is a chromogenic substrate used in biochemical assays. It is commonly employed in the detection and quantification of β-galactosidase activity, which is a widely used reporter enzyme in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
O nitrophenol, by Sangon (3 mentions)
Lactose, by Solarbio (2 mentions)
Galactose, by Solarbio (2 mentions)
Silica gel thin layer chromatography plates, by Merck Group (1 mentions)
Bl21 de3, by Takara Bio (1 mentions)
Pet28a, by Takara Bio (1 mentions)
Isopropyl β d thiogalactopyranoside, by Sangon (1 mentions)
Proteinase k, by Sangon (1 mentions)
Ampicillin, by Sangon (1 mentions)
Gentamicin, by Sangon (1 mentions)
Lysozyme, by Sangon (1 mentions)
Kanamycin, by Sangon (1 mentions)
Matchmaker gold yeast 2 hybrid system, by Takara Bio (1 mentions)
4 protocols using o nitrophenyl β d galactopyranoside
1
Heterologous Expression of β-Galactosidase
2
Escherichia coli Heterologous Protein Expression
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The E. coli strains, BL21 (DE3), and expression vector, pET28a(+), were purchased from Takara (Dalian, China). O-Nitrophenol (ONP) and o-nitrophenyl-β-D -galactopyranoside (ONPG) were purchased from Sangon Biotech (Shanghai, China). Lactose, glucose, and gaLactose used for this study were supplied by Solarbio (Beijing, China). The thin-layer chromatography (TLC) silica gel plates were purchased from Merck (Darmstadt, Germany). All other materials used were of the analytical degree.
3
Lysogeny Broth Medium Preparation
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Lysogeny broth (LB) medium (g L−1) was composed as follows: peptone 10.0 g L−1, yeast extract 5.0 g L−1, NaCl 10.0 g L−1, pH 7.2, (solid, addition of 1.5% agar), and deionized water 1000 mL, which was sterilized at 121 °C for 20 min.
Proteinase K, lysozyme, ampicillin (Amp), kanamycin (Km), gentamicin (Gm), isopropyl-β-D-thiogalactopyranoside (IPTG), o-nitrophenyl β-D-galactopyranoside (ONPG), and o-nitrophenol (ONP) were purchased from Shanghai Sangon Biotech (Sangon Biotech, Shanghai, China). GA C15:1 standard was purchased from Shanghai Tauto Biotechnology (Shanghai, China). Other chemical reagents were analytical grade.
Proteinase K, lysozyme, ampicillin (Amp), kanamycin (Km), gentamicin (Gm), isopropyl-β-D-thiogalactopyranoside (IPTG), o-nitrophenyl β-D-galactopyranoside (ONPG), and o-nitrophenol (ONP) were purchased from Shanghai Sangon Biotech (Sangon Biotech, Shanghai, China). GA C15:1 standard was purchased from Shanghai Tauto Biotechnology (Shanghai, China). Other chemical reagents were analytical grade.
4
Yeast Two-Hybrid Analysis of PC1 Interactions
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Y2H analysis was performed using the Matchmaker Gold Yeast 2-Hybrid System (Clontech, Japan) according to the manufacturer's instructions. The interactions between PC1 and CATs as well as the self-association of PC1 were verified by Y2H assays as described previously (Zhou et al. 2018) (link). The cDNAs of CatA, CatB, CatC, and PC1 were subcloned into the pGADT7 prey plasmid and individually cotransformed into the yeast strain AH109 with PC1 subcloned into the pGBKT7 bait plasmid. The primers used are listed in Supplemental Table S3. Synthetic defined (SD) medium lacking Leu and Trp was used for selection of positive transformation, while SD medium lacking Leu, Trp, His, and Ade (SD-Leu-Trp-His-Ade) was used for testing the interaction. βgalactosidase assays were performed according to the manufacturer's instructions (Clontech, Japan), using Onitrophenyl β-D-galactopyranoside (Sangon, China) as the substrate.
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