Cloning and expressing of golgi protein73 gene fragment and preparation of monoclonal antibodies against the recombinant protein / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 301-303, 2013.
Article
in Chinese
| WPRIM
| ID: wpr-318035
ABSTRACT
<p><b>OBJECTIVE</b>To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein.</p><p><b>METHODS</b>GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot was used to detect specificity of mAbs.</p><p><b>RESULTS</b>The prokaryotic plasmid expressing the recombinant protein was constructed, and the GP73 recombinant protein was expressed and purified. Five hybridoma cell lines that secreted anti-GP73 mAbs were obtained. 2 of 5 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with GP73 protein.</p><p><b>CONCLUSION</b>The GP73 recombinant protein is highly purified and has strong antigenicity. The anti-GP73 mAbs were prepared successfully.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Gene Expression
/
Cloning, Molecular
/
Allergy and Immunology
/
Hep G2 Cells
/
Genetics
/
Hybridomas
/
Liver Neoplasms
/
Membrane Proteins
/
Metabolism
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2013
Type:
Article
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