Molecular cloning of recombinant DNA : proceedings of the Miami winter symposia, January 1977 / edited by W.A. Scott, R. Werner ; sponsored by the Department of Biochemistry, University of Miami, School of Medicine, Miami, Florida.

Date:
1977
Catalogue details

Licence: Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)

Credit: Molecular cloning of recombinant DNA : proceedings of the Miami winter symposia, January 1977 / edited by W.A. Scott, R. Werner ; sponsored by the Department of Biochemistry, University of Miami, School of Medicine, Miami, Florida. Source: Wellcome Collection.

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    40 S. N. COHEN ef а/. P15a + 1 Eco RI' pSC101 CONSTRUCTION OF A PLASMID CLONING VEHICLE CONTAINING MULTIPLE SIT€S FOR INSERTIONAL INACTIVATION Cm pAC161 + - рСМ-В ; Hind III + Pst LIGATION- + TRANSFORMATION pML21 + Hin III-GENERATED Cm'' FRAGMENT OF R6-5 1. INSERTIONAL INACTIVATION AT Eco RI, Hind III, Bam I, Sal 1 SITES 2. MULTI-COPY 3. AMPLIFIABLE 4. NON-CONJUGATIVE Hae II DIGESTION, LIGATION, TRANSFORMATION, GEL FRACTIONATION 4.2 X 10® Fig. 2. Construction of pAClSl Cloning Vehicle. Construction of the plasmid is described in the text. This cloning vehicle is suitable for use with the insertional activation procedure for EcoRI, Hindlll, BamI, or Sail DNA fragments. In addition, it is a non-conjugative, multicopy cloning vector that is amplifiable in the presence of appropriate concentra¬ tions of chloramphenicol. A wide variety of other methods have been used to enable selection of plasmids carrying specific DNA fragments; in general, these involve the use of radioactively labelled DNA or RNA probes or immunological probes to identify clones carrying the desired chimeric plasmids or phage. The subcul¬ ture-cloning selection procedure employed for the cloning of histone genes from unfractionated sea urchin DNA (30) is essentially a modification of the sib-selection technique reported some years ago by Cavalli-Sforza and Lederberg (31) for the genetic identification of bacterial mutants.