Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (8): 195-202.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1361
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Received:
2020-11-05
Online:
2021-08-26
Published:
2021-09-10
Contact:
DING Yu
E-mail:980930967@qq.com;dingy@gdou.edu.cn
FAN Chen-long, DING Yu. Molecular Cloning and Functional Verification of Histone Deacetylase Gene cobB in Vibrio alginolyticus[J]. Biotechnology Bulletin, 2021, 37(8): 195-202.
Fig.2 Bioinformatics analysis of cobB sequences A:Signal peptide prediction. B:Transmembrane domain prediction.C:Protein tertiary structure prediction
Fig.4 Schematic diagram of gel electrophoresis of CobB protein expression M:Protein marker;1:negative control without IPTG;2:negative control with IPTG;3:pGEX-6p-cobB without IPTG;4:pGEX-6p-cobB with IPTG
Fig.5 Schematic diagram of gel electrophoresis of different induction time on CobB protein expression M:Protein marker;1-7:The induction time is 1 h、2 h、3 h、4 h、5 h、6 h、7 h
Fig.6 Schematic diagram of gel electrophoresis of different IPTG concentrations on CobB protein expression M:Protein marker;1-7:the IPTG concentrations are 0.1%、0.2%、0.4%、0.6%、0.8%、1% and 2%
Fig.7 Schematic diagram of gel electrophoresis of different temperatures on CobB protein expression M:Protein maker;1:28℃ bacteria protein;2:28℃ supernatant protein;3:28℃ precipitation protein;4:37℃ bacteria protein;5:37℃ supernatant protein;6:37℃ precipitation protein
Fig.8 Schematic diagram of digestion gel electrophoresis of CobB protein purification M:Protein molecular standard;1:Bacteria protein;2:Results of CobB fusion protein purification;3:Results of CobB fusion protease cleavage results
Fig.9 Western blot of CobB deacetylated GlmU protein A:Western blot of acetyllysine-specific antibodies and SDS-PAGE;B:GlmU acetylation level quantified by Western blot. 1:No CobB protein,no NAD+ group;2:CobB protein,no NAD + group;3:no CobB protein,NAD+ group;4:CobB protein,NAD+ group
Fig.10 Western blot of CobB fusion protein and CobB protein deacetylated GlmU protein A:Western blot of acetyllysine-specific antibodies and SDS-PAGE;B:GlmU acetylation level quantified by Western blot. 1:CobB fusion protein group;2:CobB protein group;3:GST protein group
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