Automated electrokinetic analysis; description and application in virology and cell biology

Area of Science:

• Biophysics
• Molecular Biology
• Analytical Chemistry

Background:

• Electrophoretic mobility (EPM) is a key parameter for characterizing charged molecules and particles.
• Automated instrumentation is crucial for precise and efficient EPM measurements.
• Bacteriophage infection significantly alters host cell surface properties.

Purpose of the Study:

• To evaluate the PenKem S3000 automated electrokinetic analyzer for measuring EPM of unbound macromolecules.
• To investigate the effects of T4 phage infection on the EPM of Escherichia coli B.
• To explore the role of phage DNA in altering host cell surface charge.

Main Methods:

• Utilized the PenKem S3000 to measure EPM of DNA, polysaccharide, and BSA in solution and bound to latex beads.
• Measured EPM of T4 phage in solution, bound to zwitterionic latex, and after sonication.
• Monitored EPM changes in E. coli B upon exposure to T4 phage and correlated with electron microscopy.

Main Results:

• The S3000 accurately determined EPM for unbound macromolecules (DNA, PCP-3, BSA).
• T4 phage infection rapidly increased E. coli B's EPM, consistent with free DNA release.
• Phage binding to zwitterionic latex suggested potential DNA release, altering EPM.

Conclusions:

• The PenKem S3000 is suitable for measuring EPM of unbound macromolecules.
• The S3000 can effectively monitor host cell surface charge dynamics during viral infection.
• Phage DNA release appears to be a significant factor in altered E. coli B surface charge post-infection.