P1 Phage and PAC cloning vector - Pac sites, Cre-loxP, and SacB lethality

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This video discusses the general biology of P1 bacteriophage based cloning vectors (pNS582 and pAd10SacBII) and P1 derived artificial chromosomes (PACs - pCYPAC1). The discussion walks you through the development of the P1 system with its use of Pac site and Cre-loxP recombination, coupled with SacB driven synthetic lethality. The main focus of the video is understanding Pac site packaging, Cre-loxP recombination and how they are used in P1 vectors (pNS582 and pAd10), along with SacB gene for counter selection. The P1 vectors are interesting in themselves because they use a cryptic origin of replication (P1 lytic origin) to increase the copy number of a vector. PACs specifically benefit from this origin, helping increase the overall vector yield in DNA extraction.

Jump to your favorite section:
00:00 P1 bacteriophage
01:19 pNS582
03:50 Cre-LoxP
05:38 Using P1 vectors
08:50 P1 Lytic origin
10:10 Disadvantages
11:58 pAd10SacBII
12:38 Using SacB for selection
14:02 PACs - pCYPAC1

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