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Identification of sequences necessary for packaging DNA into lambda phage heads
Authors:T Miwa  K Matsubara
Affiliation:Laboratory of Molecular Genetics, University of Osaka, Medical School, Kitaku, Osaka, 530 Japan
Abstract:Several species of DNA molecules are packaged into lambda phage heads if they carry the region around the cohesive end site of lambda phage (cos lambda). The minimal functional sequence around cos lambda needed for packaging was examined by cloning in pBR322. The results showed that the minimal region contained 85 bp around cos lambda; 45 bp of the left arm of lambda phage and 40 bp of the right arm. A 75-bp region located to the right of the minimal region seems to enhance packaging. A 223-bp fragment containing these regions can be used as a portable element for plasmid DNA packaging into lambda phage heads. Plasmid ppBest 322, a derivative of pBR322 carrying this portable packager and both amp and tet genes, was constructed. This plasmid is useful for cloning of large DNA fragments.
Keywords:Recombinant DNA  cosmid  M13 dideoxynucleotide sequencing  λ cohesive end  plasmid vector  gene conferring ampicillin resistance  bp  base pairs  gene conferring chloramphenicol resistance  kb  kilobase pairs  PB, PBB  see MATERIALS  MMT-PBB  METHODS Section (b)    485 bp (see Szybalski and Szybalski, 1979)  SDS  sodium dodecyl sulfate  gene conferring tetracycline resistance
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