Production and simple purification of a protein encoded by part of the gag gene of HIV-1 in the Escherichia coli HB101F+ expression system inducible by lactose and isopropyl-beta-D-thiogalactopyranoside
Jazyk angličtina Země Nizozemsko Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- chromatografie iontoměničová MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- ELISA MeSH
- Escherichia coli účinky léků genetika metabolismus MeSH
- genové produkty gag biosyntéza izolace a purifikace MeSH
- HIV-1 genetika MeSH
- isopropylthiogalaktosid farmakologie MeSH
- laktosa farmakologie MeSH
- lidé MeSH
- protilátky bakteriální izolace a purifikace MeSH
- regulace exprese virových genů účinky léků MeSH
- rekombinantní proteiny chemie izolace a purifikace MeSH
- virové geny genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- genové produkty gag MeSH
- isopropylthiogalaktosid MeSH
- laktosa MeSH
- protilátky bakteriální MeSH
- rekombinantní proteiny MeSH
The development of the Escherichia coli expression system, which was prepared by transferring the F' episome from strain 71/18 to a highly to a transformable F- strain HB101, is described. These new HB101 (F+) cells, which produced high levels of lac repressor, were capable of taking up lactose and grew under strict selection conditions. A relatively simple two-step purification of part of a protein (M(r) 27,000) encoded by the gag gene of HIV-1 in this expression system is described. The supernatant prepared by removal of cell debris was precipitated by 30% saturation of ammonium sulphate. The protein spectrum was characterized by gel electrophoresis, immunoblotting and ion-exchange titration curves. Optimum separation was achieved using a strong anion exchanger (Mono Q) at pH 8.0. The purified protein did not cross-react with antibodies to E. coli.
Citace poskytuje Crossref.org
