LOCATION AND SEQUENCE OF A PUTATIVE PROMOTER FOR THE B GENE OF BACTERIOPHAGE S13

Abstract

This chapter examines the location and sequence of putative promoter for the B gene of bacteriophage S13. The binding of Escherichia coli (E. coli) RNA polymerase to Hind II + III and Hae III restriction fragments of bacteriophage S13 has been used to localize binding sites on the phage genome. Three major binding sites were located correlating with promoter sites at the beginning of genes A and B and a site overlapping gene D and the beginning of gene E. Two less definite binding sites were also localized, one being in gene F and one at the gene G–H junction. Complexes between E. coli RNA polymerase and S13 RF DNA have also been visualized in the electron microscope. Two major locations were mapped corresponding to the beginning of genes B and D. Minor sites were also localized corresponding to position identified in the restriction fragment binding experiments. It is found that from the overall binding data, putative promoters have been placed at the start of genes A, B, and D. The site at the start of gene B has been sequenced using T4 endonuclease IV hydrolysis, a methylmethane sulfonate alkylation and alkaline hydrolysis procedure and the Maxim–Gilbert rapid sequencing method.