Bacterial Artificial Chromosome

Sp6 promoter

T7 promoter NotI , \ BamHI/NotI

1 Cloning

Sp6 promoter

Oris Bacterial Artificial Chromosome

1 Cloning

Figure 11-9 Structure of a bacterial artificial chromosome (BAC), used for cloning large fragments of donor DNA. CMP- is a selectable marker for chloramphenicol resistance. oriS, repE, parA, and parBare F genes for replication and regulation of copy number. cosN is the cos site from X phage. Hindlll and BamHI are cloning sites at which donor DNA is inserted. The two promoters are for transcribing the inserted fragment. The NotI sites are used for cutting out the inserted fragment.

Figure 11-9 Structure of a bacterial artificial chromosome (BAC), used for cloning large fragments of donor DNA. CMP- is a selectable marker for chloramphenicol resistance. oriS, repE, parA, and parBare F genes for replication and regulation of copy number. cosN is the cos site from X phage. Hindlll and BamHI are cloning sites at which donor DNA is inserted. The two promoters are for transcribing the inserted fragment. The NotI sites are used for cutting out the inserted fragment.

which enters the cell and forms a plasmid chromosome (Figure 11-10a). When phages are used, the recombinant molecule is combined with the phage head and tail proteins. These engineered phages are then mixed with the bacteria, and they inject their DNA cargo into the bacterial cells. Whether the result of injection will be the introduction of a new recombinant plasmid (Figure 11-10b) or the production of progeny phages carrying the recombinant DNA molecule (Figure 11-10c) depends on the vector system. If the latter, the resulting free phage particles then infect nearby bacteria. When X phage is used, through repeated rounds of reinfection, a plaque full of phage particles, each containing a copy of the original recombinant X chromosome, forms from each initial bacterium that was infected.

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