A general method for inserting specific DNA sequences into cloning vehicles |
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Authors: | Chander P Bahl Kenneth J Marians Ray Wu |
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Institution: | Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853 U.S.A.;Division of Biological Sciences, National Research Council of Canada, Ottawa, Ontario Canada |
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Abstract: | A general method has been developed to introduce any double-stranded DNA molecule into cloning vehicles at different restriction endonuclease sites. In this method a chemically synthesized decadeoxyribonucleotide duplex, containing a specific restriction endonuclease sequence, is joinlex DNA is cut by the same restriction endonuclease to generate the cohesive ends. It is then inserted into the restriction endonuclease cleavage site of the cloning vehicle. To demonstrate the feasibility of this new method, we have inserted separately the synthetic lac operator DNA at the Bam I and HindIII cleavage sites of the plasmid pMB9 DNA. |
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Keywords: | Synthetic decadeoxyribonucleotides linker sequences lac DNA plasmids restriction endonucleases cloned DNA |
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