About: Expression and Purification of Human Keratinocyte Growth Factor 2 by Fusion with SUMO

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An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Expression and Purification of Human Keratinocyte Growth Factor 2 by Fusion with SUMO
Creator	Wu, Xiaoping Su, Zhijian Ae, Huang Ae, Jian Ae, Nie Ae, Tan Ae, Xiao Changjun, A Feng, Wenke Huang, Zhifeng Li, Ae Yadong, A Yan, Qiuxia Yanfang, A Yecheng, A Yi, A Zeng, Yaoying
source	Medline; PMC
abstract	Small ubiquitin-related modifier (SUMO) fusion system has been shown to be efficient for enhancing expression and preventing degradation of the target protein. We showed herein that SUMO fusion to human keratinocyte growth factor 2 (hKGF-2) gene was feasible and it significantly enhanced protein expression and its efficiency. The fusion DNA fragment composed of SUMO gene, which was fused to hexahistidine tag, and hKGF-2 gene was amplified by PCR and inserted into the expression vector pET28a to construct the recombinant plasmid, pET28a-SUMO-hKGF-2. The plasmid was then transformed into Escherichia coli Rosetta(TM)2(DE3), and the recombinant fusion protein SUMO-hKGF-2 was expressed at 30°C for 6 h, with the induction of IPTG at the final concentration of 0.4 mM. The expression level of the fusion protein was up to 30% of the total cellular protein. The fusion protein was purified by Ni-NTA affinity chromatography. After desalting by Sephadex G-25 size exclusion chromatography, the hexahistidine-SUMO-hKGF-2 was digested by SUMO proteases. The recombinant hKGF-2 was purified again with Ni-NTA column and the purity was about 95% with a total yield of 13.9 mg/l culture. The result of mitogenicity assay suggests that the recombinant hKGF-2 can significantly promote the proliferation of normal rat kidney epithelial (NRK-52E) cells.
has issue date	2008-12-18(xsd:dateTime)
bibo:doi	10.1007/s12033-008-9135-7
bibo:pmid	19104760
has license	no-cc
sha1sum (hex)	e9c1b8e260f20d49172aef5873c4452a219b699c
schema:url	https://doi.org/10.1007/s12033-008-9135-7
resource representing a document's title	Expression and Purification of Human Keratinocyte Growth Factor 2 by Fusion with SUMO
has PubMed Central identifier	PMC7090735
has PubMed identifier	19104760
schema:publication	Mol Biotechnol
resource representing a document's body	covid:e9c1b8e260f20d49172aef5873c4452a219b699c#body_text
is schema:about of	named entity 'SUMO' named entity 'FUSION' named entity 'FUSION' named entity 'PREVENTING' named entity 'KERATINOCYTE GROWTH FACTOR 2' covid:arg/e9c1b8e260f20d49172aef5873c4452a219b699c named entity 'PURIFICATION' named entity 'EXPRESSION' named entity 'HUMAN' named entity 'SUMO' named entity 'MODIFIER' named entity 'ENHANCING' named entity 'DEGRADATION' named entity 'TARGET PROTEIN' named entity 'EXPRESSION' named entity 'SMALL' named entity 'EFFICIENT' named entity 'UBIQUITIN-RELATED' named entity 'SYSTEM' named entity 'molecular weight' named entity 'Carlsbad, California, USA' named entity 'total protein' named entity 'plasmid' named entity 'fusion protein' named entity 'NRK' named entity 'protein' named entity 'imidazole' named entity 'fusion protein' named entity 'FGFR2' named entity 'venous ulcers' named entity 'fusion gene' named entity 'IPTG' named entity 'KGF' named entity 'therapeutic protein' named entity 'genome' named entity 'pH 8' named entity 'E. coli' named entity 'Ni-NTA' named entity 'FGF' named entity 'PCR' named entity 'epidermal' named entity 'SUMO' named entity 'IPTG' named entity 'pH 8' named entity 'FGF receptor' named entity 'codons' named entity 'ulcer' named entity 'solubility' named entity 'fusion gene' named entity 'KGF' named entity 'regulation of gene expression' named entity 'fusion protein' named entity 'SUMO' named entity 'codons' named entity 'oral mucositis' named entity 'hydrolase' named entity 'primers' named entity 'Western blotting' named entity 'kanamycin' named entity 'plasmid' named entity 'Invitrogen' named entity 'heparin' named entity 'peptides' named entity '19.3' named entity 'protein expression'

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