A quantitative theory of transfection in B. subtilis.

ABSTRACT

A theory of transfection is developed which describes three different types of experiments in transfection The concentration dependence of transfection, transfection with marker rescue, and the mapping function in transfection crosses. The theory is applicable to transfection systems exhibiting quadratic or higher order dose response (APP1, AP50, SP82). It pictures the essential process in transfection as follows transfecting DNA molecules, having suffered inactivating events during uptake or intracellularly, have to recombine prior to replication under elimination of these lesions. The probability for recombination, successful in this sense, is calculated as a function of the number of DNA molecules within the competent cell, the mean number of inactivating events per DNA molecule, and the crossover probability per nucleotide. Under the assumption of random distribution of inactivating events over the population of DNA molecules and homogeneous crossover probabilities the theory explains on a quantitative basis a number of experimental observations in transfection, as for instance the relative efficiencies of different helper phage in transfection with marker rescue, the third order concentration dependence in SP50 transfection, and the high recombination frequencies observed in transfection crosses.