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Published January 1994 | metadata_only
Journal Article

A new bacteriophage P1-derived vector for the propagation of large human DNA fragments


We have designed a P1 vector (pCYPAC−1) for the introduction of recombinant DNA into E. coli using electroporation procedures. The new cloning system, P1−derived arteficial chromosomes (PACs), was used to establish an initial 15,000 clone library with an average insert size of 130−150 kilobase pairs (kb). No chimaerism has been observed in 34 clones, by fluorescence in situ hybridization. Similarly, no insert instability has been observed after extended culturing, for 20 clones. We conclude that the PAC cloning system will be useful in the mapping and detailed analysis of complex genomes.

Additional Information

© 1994 Nature Publishing Group. Received 28 September; accepted 18 October. P.A.I. and C. T.A contributed equally to this work. We thank Joel Jessee (BRL/Life Technologies, Inc.) Nat Sternberg for their help and contribution to this work. P.A.I. was partially supported by a grant from the Muscular Dystrophy Association, U.S.A., and by a Unesco/TWAS short term fellowship in the Human Genome Project. P.M.K. was supported in part by the FWF grant J0720-MED. Work performed at the Lawrence Livermore National Laboratory under the auspices of the U.S. Department of Energy under contract No. W-7405-ENG-48.

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August 20, 2023
August 20, 2023