Volume 3

Gene expression / Benjamin Lewin.

  • Lewin, Benjamin.
Date:
1974-1977
Catalogue details

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

Credit: Gene expression / Benjamin Lewin. Source: Wellcome Collection.

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    Eco RI cleavage sites 1 2 3 4 5 A BCD E F isolate mutant lacking sites 4 and 5 > f I Ä B C D-E-F cleave with Eco RI nuclease \ t isolate Agt—0 lacking B and C y A D-E-F Figure 3.24: Eco RI cleavage of \ DNA and the preparation of Xgt -0. Cleavage sites are indicated by the shaded arrows; the fragments released are indicated by letters A-F. Upon cleavage with Eco RI, these release the terminal fragments A and D-E-F, which can anneal with a foreign DNA to form a viable phage. Cameron et al. (1975) have used this technique to clone X DNA carrying the E.coli ligase gene; and Kramer, Cameron and Davis (1976) have cloned yeast genes inserted in A. DNA. Replication of Composite Plasmids In addition to rearranging the sequences of existing plasmids and inserting foreign DNA into them, it is possible to use the Eco RI cleavage tech nique to construct new plasmids that contain components from naturally occurring but unrelated plasmids that usually do not recombine. Timmis, Cabello and Cohen (1974) have constructed the interesting composite plasmid pSC101-Col El by transforming E.coli cells with DNA of the two plasmids that was cleaved with Eco RI, mixed, and ligated: the composite