Experiments on expression of cloned genes in E. coli using λ phage PL promoter
The λ phage PL promoter is a potent promoter controlled by a temperature-sensitive repressor (cIts857), which can repress transcription of the PL promoter at l
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Experiments on secretion and expression of exogenous proteins with alkaline phosphatase promoter and signaling sequences
This experiment describes the process of expressing exogenous proteins in Escherichia coli using the alkaline phosphatase promoter (phoA) and signaling sequence
Lysogenic infection assay on agar plates
Fusion proteins encoded by λ phage recombinants can be prepared by lysogenizing colonies of E. coli strain Y1089. However, preparation of lysogenic bacteria fr
Experimental preparation of λ phage particles from large-scale lysates
λ phage prepared from mass cultures can be recovered from the lysate by polyethylene glycolysis (PEG) precipitation in the presence of high salt. The residual
Purification of expressed proteins from inclusion bodies
This experiment was mainly used to obtain soluble active proteins.
Lipofection-mediated DNA transfection assay
The following method is a modification of one studied by Mark Evans (Alexion Pharmaceuticals, New Haven, Connecticut). This experiment was derived from the next
Experimental extraction of λ phage DNA from mass cultures using proteinase K and SDS
DNA can be well isolated from large-scale preparations of λ phage by digestion with an effective protease (e.g., proteinase K) and subsequent phenol:chloroform
Calcium phosphate-mediated plasmid DNA transfection in eukaryotic cells
The calcium phosphate-mediated plasmid DNA transfection of adherent cells described in this method is an improvement on the method established by Jordan et al.
Experiments on the purification of proteins with histidine tags by solidified Ni2+ absorption chromatography
Polyhistidine binds to a variety of transition metals and transition metal chelates, so proteins with exposed His-6 bind to cured Ni2+ resins, and target protei