We describe in vitro conditions for packaging of exogenous DNA of Salmonella phage P22 which has terminally redundant, circularly permuted DNA. The method is a modification of the Kaiser-Masuda procedure. The most important aspect is to prepare all components (proheads, enzymes and concatemeric DNA) in end− cells. The influence of several factors such as DNA- and Mg2+ concentration and kinetics has been investigated.
BotsteinD.,
WaddellC. H.,
KingJ.1973; Mechanism of head assembly and DNA encapsulation in Salmonella phage P22. I. Genes, proteins, structures and DNA maturation. Journal of Molecular Biology 80:669–695
ChanR. K.,
BotsteinD.1972; Genetics of bacteriophage P22. I. Isolation of prophage deletions which affect immunity to superinfection. Virology 49:257–267
EarnshawW.,
KingJ.1978; Structure of P22 coat protein aggregates formed in the absence of scaffolding protein. Journal of Molecular Biology 126:721–747
KaiserD.,
MasudaT.1973; In vitro assembly of bacteriophage lambda heads. Proceedings of the National Academy of Sciences of the United States of America 70:260–264
RajA. S.,
RajA. Y.,
SchmiegerH.1974; Phage genes involved in the formation of generalized transducing particles in Salmonella phage P22. Molecular and General Genetics 135:175–184
SchmiegerH.1968; Die molekulare Struktur transduzierender Partikel beim Salmonella Phagen P22.I. Dichtegradienten–Untersuchungen an intakten Phagen. Molecular and General Genetics 102:336–347
SchumannW.,
BadeE. G.1977; A Salmonella typhimurium endonuclease that converts native DNA to fragments of about 8 x 1o5 daltons. Journal of General Microbiology 101:319–325
StreisingerG.,
EmrichJ.,
StahlM. M.1967; Chromosome structure in phage T4. III. Terminal redundancy and length determination. Proceedings of the National Academy of Sciences of the United States of America 57:292–295
SusskindM. M.,
WrightA.,
BotsteinD.1971; Superinfection exclusion by P22 prophage in lysogens of Salmonella typhimurium. II. Genetic evidence for two exclusion systems. Virology 45:638–652