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Construction of a prokaryotic expression vector for apoptin and preparation of polyclonal antibody of apoptin / 南方医科大学学报
Journal of Southern Medical University ; (12): 1405-1407, 2009.
Article in Chinese | WPRIM | ID: wpr-268747
ABSTRACT
<p><b>OBJECTIVE</b>To construct a prokaryotic expression vector for apoptin and prepare polyclonal antibody of apoptin.</p><p><b>METHODS</b>Apoptin gene amplified from pGEM-T/Apoptin plasmid by PCR was cloned into pET-28a (+). E.coli BL21 (DE3) was transformed by the recombinant plasmid, and apoptin protein expression induced by IPTG was analyzed by SDS-PAGE. BALB/c mice were immunized with the protein and the titer of the antibody was determined using indirect enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Apoptin gene was successfully cloned into pET-28a (+), and the expression of a protein with relative molecular mass of about 17 000 was identified by SDA-PAGE. After 5 immunizations of the mice with the protein, the blood antibody titer reached 15x10(5).</p><p><b>CONCLUSION</b>The prokaryotic expression vector for apoptin is successfully constructed and the polyclonal antibody of apoptin is obtained, which allows further functional study of apoptin.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Blood / Gene Expression / Genome / Cloning, Molecular / Capsid Proteins / Allergy and Immunology / Escherichia coli / Genetic Vectors / Genetics Limits: Animals Language: Chinese Journal: Journal of Southern Medical University Year: 2009 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Plasmids / Blood / Gene Expression / Genome / Cloning, Molecular / Capsid Proteins / Allergy and Immunology / Escherichia coli / Genetic Vectors / Genetics Limits: Animals Language: Chinese Journal: Journal of Southern Medical University Year: 2009 Type: Article