Xi bao yu fen zi mian yi xue za zhi, 2007-01, Vol.23 (1), p.25
2007
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Details
- Autor(en) / Beteiligte
- Titel
- Construction of the expression vector pET32a/His MBL-CLR and its expression in E.coli
- Ist Teil von
- Xi bao yu fen zi mian yi xue za zhi, 2007-01, Vol.23 (1), p.25
- Ort / Verlag
- China
- Erscheinungsjahr
- 2007
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- To construct pET32a/His MBL-CLR recombinant prokaryotic expression plasmid and to express mannan-binding lectin-CLR (MBL-CLR) protein in E.coli The human MBL-CLR gene was amplified by PCR from pGEM-MBL plasmid, and was inserted into prokaryotic expression vector pET32a. After identified by restriction mapping and sequencing, the recombinant plasmid pET32a/His MBL-CLR was transformed into E.coli BL21 (DE3) cells. The expressed product was purified by Immobilized Metal Affinity Chromatography (IMAC) and identified by SDS-PAGE, Western blot and indirect enzyme-linked immunosorbent assay (ELISA) using the antibody from BALB/c mice immunized with the recombinant human MBL protein. The cDNA fragment of 180 bp was amplified from pGEM-MBL plasmid and the recombinant expression vector pET32/His MBL-CLR was constructed. The recombinant plasmid was consistent with those expected by restriction maps and sequence. Three components of relative molecular mass 30,000, 60,000 and 120,000 in the purified recombinant product we
- Sprache
- Chinesisch
- Identifikatoren
- ISSN: 1007-8738Titel-ID: cdi_pubmed_primary_17210100
- Format
- –
- Schlagworte
- Animals, Blotting, Western, Chromatography, Affinity, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Escherichia coli - genetics, Escherichia coli - metabolism, Genetic Vectors - genetics, Humans, Mannose-Binding Lectin - genetics, Mannose-Binding Lectin - metabolism, Mice, Mice, Inbred BALB C, Polymerase Chain Reaction, Recombinant Fusion Proteins - genetics, Recombinant Fusion Proteins - immunology, Recombinant Fusion Proteins - metabolism