Stability of steady states in nucleic acid poly[d(A-T)] synthesis and the stirred flow reactor

✍ Scribed by Friedemann W. Schneider

• Publisher: Wiley (John Wiley & Sons)
• Year: 1976
• Tongue: English
• Weight: 782 KB
• Volume: 15
• Category: Article
• ISSN: 0006-3525
• DOI: 10.1002/bip.1976.360150102

✦ Synopsis

Abstract

The template directed synthesis of poly[d(A‐T)] from the nucleoside triphosphates in the presence of DNA polymerase I is carried out continuously in a stirred flow reactor for the first time. The initial objective is to test the kinetic stability of the established steady states at various flow rates. Graphical analysis predicts instable steady states for certain high flow rates. As a consequence of instabilities multiple steady states and steady‐state hysteresis may occur. Steady‐state hysteresis has now been found experimentally. For a different enzyme fraction of low exonuclease activity we found the steady‐state absorbance at 260 nm to be almost invariant with flow rate at high enzyme concentrations even if the flow rate was increased by a large factor. We call this phenomenon kinetic buffering. Relaxation of a large flow perturbation approaches the steady state in a sigmoidal fashion. Concentration oscillations at 260 nm occurred in one experiment using an enzyme fraction of low exonuclease activity after perturbing the steady state by monomer (dATP). Advantages of the stirred flow reactor method over serial transfer are discussed.