Increased protein yields from Escherichia coli using pressure-cycling technology.
Sample preparation is critical to the success of two-dimensional gel electrophoresis and other analytical methods. Pressure-cycling technology (PCT) uses alternating cycles of high and low pressure to induce cell lysis. Cell suspensions were placed in PULSE Tubes and subjected to alternating cycles of high and low pressure in a Barocycler instrument. each cycle consisted of 20 sec at 35,000 psi followed by 20 sec at ambient pressure. For the bacterium Escherichia coli, PCT extracted 14.2% more total protein than was extracted using a standard bead mill. Image analysis of two-dimensional gels revealed 801 protein spots in the PCT lysate, compared to 760 protein spots in the bead mill lysate.
Smejkal
Gary B
GB
Pressure BioSciences, Proteomics Laboratory, 6 Gill Street, Woburn, MA 01801, USA. gsmejkal@pressurebiosciences.com
Robinson
Myra H
MH
Lawrence
Nathan P
NP
Tao
Feng
F
Saravis
Calvin A
CA
Schumacher
Richard T
RT
D001709:Biotechnology / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D007091:Image Processing, Computer-Assisted; D009991:Oscillometry; D011312:Pressure; D040901:Proteomics / Q000379:methods
D029968:Escherichia coli Proteins