Molecular Cloning: Ch. 15. Expression of cloned genes in Escherichia coli |
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Page 15-19
... mutations in the lon gene , which encodes the Lon ( or La ) protease , an ATP - dependent endo- protease . This protease , working in conjunction with several heat shock proteins , degrades unfolded and abnormal ( foreign ) proteins in ...
... mutations in the lon gene , which encodes the Lon ( or La ) protease , an ATP - dependent endo- protease . This protease , working in conjunction with several heat shock proteins , degrades unfolded and abnormal ( foreign ) proteins in ...
Page 8
... mutations that may be used for complementation with yeast TRP1 , LEU2 , and HIS3 wild - type mark- ers . Note that the Tn5 transposon confers kanamycin resistance . A strain that will support growth of vectors carrying amber mutations ...
... mutations that may be used for complementation with yeast TRP1 , LEU2 , and HIS3 wild - type mark- ers . Note that the Tn5 transposon confers kanamycin resistance . A strain that will support growth of vectors carrying amber mutations ...
Page 5
... mutations because of wobble in the third position of the codon . At one time , it was mandatory for bacteriophage λ vectors to carry amber mutations in genes encoding coat proteins . It was believed that such mutations might reduce the ...
... mutations because of wobble in the third position of the codon . At one time , it was mandatory for bacteriophage λ vectors to carry amber mutations in genes encoding coat proteins . It was believed that such mutations might reduce the ...
Contents
Chapter | 15-3 |
Expression of Cloned Genes in Escherichia coli 15 | 15-12 |
INFORMATION PANELS | 15-55 |
Copyright | |
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activity affinity aliquots ampicillin antibody Appendix assay B-galactosidase bacterial bacteriophage bacteriophage T7 baculovirus binding Biochem Biol cDNA cell lines cell lysate centrifugation Chapter chloroquine chromatography cleavage cloning codon concentration coprecipitate DEAE-dextran dishes dithiothreitol DNase EDTA efficiency electrophoresis elution encoding enzyme Escherichia coli expression system expression vector fluorescent foreign protein fusion protein gene genomic glycerol green fluorescent protein growth growth medium H₂O hybridization inclusion bodies Incubate the cultures induced interactions lacZ luciferase lysis lysozyme mammalian cells medium membrane method mg/ml minutes at 4°C minutes at room mutations NaCl Natl Nucleic Acids pellet peptide periplasmic phosphate plasmid plasmid DNA plates promoter protease protein of interest Protocol purification reaction recombinant refolding repressor requires the reagents resin Resuspend RNA polymerase room temperature rotor SDS gel-loading buffer SDS-polyacrylamide gel sequence solubilization Sorvall ẞ-galactosidase Step sterile stock solutions strain supernatant target protein tein transcription transfection Tris-Cl pH volume wash yeast