Cloning the complete rIIB gene of bacteriophage T4 and some observations concerning its middle promoters

Abstract

We cloned the intact T4 rHB gene by joining plasmids carrying gene fragments. rIlB was expressed at a low level under control of the lac promoter, and the clone complemented rIIB mutants. We suspect that earlier attempts to clone the intact gene were unsuccessful because of transcription from T4 middle-mode promoters. These promoters are silent early in infection but are recognized when resident on a plasmid in an uninfected cell.

Wild-type bacteriophage T4 is restricted by the lambda rex genes

“…From BamHI clockwise to EcoRI, it was derived from pBR322. (b) prex2l was constructed by cloning the rex genes into pRB6 (36). (c) prex20 was constructed by cloning the rex genes into pACYC184.…”

“…pGW7 restricts rII mutants (but not wild-type T4) at low temperatures, but cells containing it grow too poorly at high temperatures to be tested. pRB6 is an adaptation of pGL101 (20) made by inserting a BamHI dodecamer linker into the PvuII site (36). pACYC184 (5) was provided by Mike Kahn.…”

The rIIA gene of bacteriophage T4. I. Its DNA sequence and discovery of a new open reading frame between genes 60 and rIIA.

“…The rIIB gene has been sequenced (PRIBNOW et al 198 1 ; HUANG 1986) and has been the subject of recent studies on translation (SHINEDLING et al 1987a,b) and transcription (SHINEDLING, WALKER and GOLD 1986). The wealth of genetic information concerning the Biotechnologie, I.N.R.A.…”