Pian13 From Pian7, Origin of Replication, pUC Polycloning, Plasmid Vector, Recombinant Plasmid, Amber Mutations, Essential Genes

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ΠAN13 (drived from ΠAN7)

ΠAN13 was derived from ΠAN7, which in its turn was derived from ΠVX. It contains a pBR322 origin of replication, pUC polycloning site and a supF tRNA gene. ΠAN13 has been used for recombination screening.

Probe sequence is cloned in the ΠAN13 plasmid vector carrying supF and the recombinant plasmid is introduced into E.coil. Libraries constructed in phage λ vectors that carry several amber mutations in essential genes are also grown in bacteria containing recombinant ΠAN13 plasmid.

If there are homologous sequences between plasmid and λ vectors, the whole plasmid is transferred to λ vectors by recombination, so that in the cointegrants, supF of πAN13 suppresses amber mutation in one or more essential genes carried by the phage thus permitting plaque formation in non-suppressing hosts.

Thus the genes carrying sequences homologous to the probe carried by πANl3 (e.g. multi gene families, genomic sequence corresponding to eDNA, etc.) can be isolated.

Plasmid PiAN13 (carrying supF) and its characteristic features.

Plasmid PiAN13 and its characteristic features

1. 698 Mael	2. 784 Hgl EII	3. 830 FnuDII
4. 853 MboII	5. FspII 46	6. Eae, GdhI85
7. Aval 91	8. SfaNI 152	9. FunDII 168
10. MseII 187 Xmnl 194 polycloning site	11. SlaNI 291	12. BstlNI 351
13. BstlNI 364	14. HaeII 447	15. 478 Ddel
16. 517 Bap 1266, ApeLI, HglAI	17. ori	18. AN13 (0.895 kb)

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