Preparation and sequencing of the cloacin fragment of Streptomyces aureofaciens 16S RNA |
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| Authors: | I Janda K Mikulík |
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| Affiliation: | 1. College of Sciences and Mathematics and Molecular Biosciences Program, Arkansas State University, Jonesboro, Arkansas, United States;2. Department of Biological Sciences, Arkansas State University, Jonesboro, Arkansas, United States;3. Audubon Delta, National Audubon Society, Baton Rouge, Louisiana, United States;4. United States Department of Agriculture, Natural Resources Conservation Service, Addis, Louisiana, United States;1. Department of Genetic Engineering and Biotechnology, Molecular Biology and Protein Science Laboratory (MBPSL), University of Rajshahi, Rajshahi 6205, Bangladesh;2. Bangladesh Sericulture Development Board, Rajshahi, Bangladesh;3. Institute for Developing Science and Health Initiatives (ideSHi), 167/24, Blue moon Tower, 11th Floor ECB Chattar, Shagufta New Rd, Dhaka 1216, Bangladesh;1. Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843, United States;2. Texas A&M AgriLife Research and Extension Center, Uvalde, TX 78801, United States |
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| Abstract: | A fast method for isolation of a 3'-terminal fragment of Streptomyces aureofaciens 16S RNA was developed. The procedure involves reaction of 70S ribosomes with cloacin DF13 and subsequent fractionation of the reaction mixture by polyacrylamide gel electrophoresis. The cloacin fragment was eluted from the gel and used directly for 3'-end labeling with cytidine-3',5'-[5'-32P]bisphosphate. The labeled RNA fragment was sequenced by the enzymatic method. It consists of 50 nucleotides and has the sequence 5'-GUCGUAACAAGGUAACCGUACCGGA-AGGUGCGGUUGGAUCACCUCCUUUCOH. The differences from the E. coli and Bacillus sequences and their possible influence on the rate and specificity of polypeptide synthesis are discussed. |
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