A simplified procedure for the rapid identification of recombinant pAT153 plasmids in Escherichia coli HB101 cells |
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| Authors: | M Dion J Yelle C Hamelin |
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| Institution: | 1. KU Leuven – University of Leuven, Department of Neurosciences, Leuven Brain Institute (LBI), Leuven, Belgium;2. Laboratory of Neurobiology, VIB Center for Brain & Disease Research, Leuven, Belgium;3. Department of Human Genetics, University Hospitals Leuven, Leuven, Belgium;4. Department of Neurology, University Hospitals Leuven, Leuven, Belgium;1. Department of Chemistry, Faculty of Mathematics and Natural Sciences, University of Mataram, Jl. Majapahit No. 62, Mataram, 83125, West Nusa Tenggara, Indonesia;2. Laboratory of Analytical Chemistry, Faculty of Mathematics and Natural Sciences, University of Mataram, Jl. Majapahit No. 62, Mataram, 83125, West Nusa Tenggara, Indonesia;3. Department of Pharmacy, Faculty of Medical, University of Mataram, Jl. Majapahit No. 62, Mataram, 83125, West Nusa Tenggara, Indonesia;4. Department of Physics, Faculty of Mathematics and Natural Sciences, University of Mataram, Jl. Majapahit No. 62, Mataram, 83125, West Nusa Tenggara, Indonesia;5. Department of Chemistry, Faculty of Science and Data Analytics, Institut Teknologi Sepuluh Nopember, Jl. Arief Rahman Hakim, Sukolilo, Surabaya 60111, Indonesia |
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| Abstract: | Insertion of foreign DNA into the unique HindIII site of the high copy number plasmid pAT153 reduces but does not completely abolish the resistance of Escherichia coli HB101 cells to tetracycline. Recombinant DNA-containing colonies could then be phenotypically differentiated from non-recombinant ones by their smaller size on nutrient agar plates with ampicillin and tetracycline at a final concentration of 50 and 4 micrograms/ml, respectively. A wide variety of human cytomegalovirus DNA fragments have been found in pAT153 molecules propagated by the ampicillin-resistant tetracycline-sensitive bacteria selected. |
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