Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (6): 1173-1180.doi: 10.16420/j.issn.0513-353x.2020-0557
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YU Qinhan1, JIAO Shuzhen1, WU Nan1, ZHANG Ningbo2, XU Weirong2,*()
Received:
2020-12-21
Revised:
2021-03-21
Online:
2021-06-25
Published:
2021-07-08
Contact:
XU Weirong
E-mail:xuwr@nxu.edu.cn
CLC Number:
YU Qinhan, JIAO Shuzhen, WU Nan, ZHANG Ningbo, XU Weirong. Molecular Cloning,Expression and Polyclonal Antibody Preparation of E3 Ubiquitin Ligase Gene HOS1 from Vitis vinifera[J]. Acta Horticulturae Sinica, 2021, 48(6): 1173-1180.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2020-0557
Fig. 2 Prediction of VvHOS1 protein domain Zf-C3H4_3:RING finger domain of C3H4 type;ELYS:Embryonic large molecule derived from yolk sac;Line:Region used for prokaryotic expression.
Fig. 4 Analysis of prokaryotic expression of pET28a-VvHOS11-320 A:The construction process of fusion protein prokaryotic vector pET28a-VvHOS11-320;B:Prokaryotic protein induced expression of pET28a-VvHOS11-320. M:Protein molecular weight standard;CK:Induced expression products of pET28a empty vector.
Fig. 5 Assay of solubility of recombinant protein 1:0.4 mg ∙ mL -1 BSA;M:Protein molecular weight standard;2:Lysis supernatant;3:The supernatant(dissolved by 2 mol · L -1urea);4,5:Inclusion bodies of 2-fold and 10-fold diluents (dissolved by 8 mol · L -1 urea);6:Induced expression of empty vector pET28a.
Fig. 6 SDS-PAGE analysis of VvHOS11-320 50 kD purified fusion protein 1,2:0.4 mg ∙ mL -1 BSA;M:Protein molecular weight standard;3,4:10-fold and 20-fold dilution of inclusions (6 mol · L -1 guanidine hydrochloride dissolving inclusion bodies).
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