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By Ray Wu, Lawrence Grossman, Kivie Moldave
FROM THE PREFACE: interesting new advancements in recombinant DNA examine let the isolation and amplification of particular genes or DNA segments from nearly any residing organism. those new advancements have revolutionized our ways to fixing complicated organic difficulties and feature spread out new probabilities for generating new and higher items within the components of wellbeing and fitness, agriculture, and industry.Volumes a hundred and one hundred and one complement Volumes sixty five and sixty eight of tools in Enzymology. over the last 3 years, many new or stronger tools on recombinant DNA or nucleic acids have seemed, and they're integrated in those volumes. quantity a hundred covers using enzymes in recombinant DNA examine, enzymes affecting the gross morphology of DNA, proteins with really good features performing at particular loci, new equipment for DNA isolation, hybridization, and cloning, analytical tools for gene items, and mutagenesis: in vitro and in vivo. quantity one hundred and one contains sections on new vectors for cloning genes, cloning of genes into yeast cells, and structures for tracking cloned gene expression.
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Additional resources for Recombinant DNA, Part B
Sample text
Ehrlich, and M. Ehlich, Nucleic Acids Res. 10, 1579 (1982). u H. Youssoufian and C. Mulder, J. Mol. Biol. 150, 133 (1981). 16 ENZYMES IN RECOMBINANT D N A [1] When 30 type II restriction endonucleases were separately incubated with Xanthomonas oryzae phage XP12 DNA, all cytosine residues of which are modified to 5-methylcytosine, only TaqI cleaved efficiently. When bacteriophage T4 DNA, which contains only 5-hydroxymethylcytosine, but not cytosine, was tested, again only TaqI cleaved, although inefficiently.
Neuendorf, in "Gene Amplification and Analysis," Vol. I: "Restriction Endonucleases" (J. G. ), p. 101. Elsevier/North-Holland, Amsterdam, 1981. 31 p. L. Molloy, and R. H. Symons, Nucleic Acids Res. 8, 2939 (1980). 32 j. LeBon, C. Kado, L. Rosenthal, and J. G. Chirikjian, Proc. Natl. Acad. Sci. A. 74, 542 (1977). 5 -- H. Belle Isle, unpublished results, 1981. b Activity was measured by incubation of 1/~g of the specified DNA with various amounts of the respective restriction endonucleases under appropriate standard reaction conditions.
When bacteriophage T4 DNA, which contains only 5-hydroxymethylcytosine, but not cytosine, was tested, again only TaqI cleaved, although inefficiently. The complete substitution of thymine residues with 5-hydroxymethyluracil in the genome of Bacillus subtilis phages SP01 and PBS1 either had no effect or for, some of the restriction enzymes, only reduced cleavage efficiency. The substitution of thymine by phosphogluconated or glucosylated 5-(4',5'-dihydroxy)pentyluracil in B. subtilis phage SP15 DNA precluded cleaving by most of the restriction endonucleases tested.