Overproduction and purification of the products of bacteriophage T3 genes 18 and 19, two genes involved in DNA packaging.
The products of gene 18 (gp18) and gene 19 (gp19) of bacteriophage T3 are noncapsid proteins involved in DNA packaging. A restriction fragment containing gene 18 or 19 was cloned into the plasmid vector pNT45 under the control of the inducible leftward promoter (PL) of phage lambda. Induction of transcription of gene 18 or 19 by derepression of the PL promoter led to the synthesis of a high level of gp18 or gp19. By using complementation of T3 DNA packaging in vitro as an assay, gp18 and gp19 were purified to near homogeneity. The overall yields of gp18 and gp19 were 1.4 mg and 0.35 mg, respectively, from 1 g wet wt cells. Addition of gp18 to the in vitro DNA packaging system resulted in increased phage production with increasing amounts of gp18 until 10% of the DNA was packaged into infectious phage particles. In contrast, addition of gp19 to the packaging system initially caused an increase in phage production, but increasing amounts of gp19 inhibited DNA packaging.
D003001:Cloning, Molecular; D004279:DNA, Viral / Q000378:metabolism*; D005814:Genes, Viral; D010957:Plasmids; D013604:T-Phages / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D004279:DNA, Viral; D014764:Viral Proteins