Recombination of a eukaryotic DNA in bacteria

Abstract

Single, 824 bp repeating units of xenopus laevis oocyte-type 5S DNA were inserted into the recombination vectors, lambda rva and lambda rvb. When the inserts had the same orientation with respect to the lambda chromosomes. Spi- imm434 recombinants were recovered by selection on a P2, lambda double lysogenic host. Because of the structure of the vectors, the crossover point in each recombinant must lie completely within the 5S DNA insert. The physical characteristics of these recombinants were determined by examination of restriction enzyme digests. By use of RecA mutant hosts and the Red- vector, lambda rvc, recombination frequencies were measured separately for the bacterial and phage systems. Some of the recombination events resulted in 5S DNA inserts of altered length due to unequal crossovers within repeated sequences in the 5S DNA spacer. The occurrence of just such events in frog 5S DNA had been predicted, based on the structure of 5S DNA and evolutionary considerations.