The Structure and Function of the DNA from Bacteriophage Lambda

The position and orientation of genes in lambda and lambda dg DNA are described. The position of six genes located in the right half of isolated lambda DNA was found to be -(N, i (λ))--O-P---Q-R-(right end of DNA), which is their order on the genetic map of the vegetative phage. The order of the three genes of the galactose operon (k, t, and e) located in the left half of lambda dg DNA was found to be (left end of DNA)----k-t-e-, consistent with Campbell's model (5) for the formation of this variant. Gene orientation, defined as the direction of transcription along the DNA, is inferred to be from right to left for the galactose operon in lambda dg DNA. The strand of lambda DNA which functions as template in transcription of N, an "early" gene required for normal replication of lambda DNA, was determined as a first step in ascertaining the orientation of this gene. The method includes isolation of each strand, formation of each of two heteroduplex molecules consisting of one strand from wild-type and one from an N mutant) and comparison of their N activities. The second step, which consists of ascertaining the 5'-to-3' direction of each strand, is discussed, as is a determination of the orientation of gene R.