Effective method for obtaining long nucleotide chains on partially complementary templates. Processed bovine gamma-interferon gene obtained from human gamma-interferon gene |
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| Authors: | V M Rostapshov I P Chernov T L Azhikina A M Borodin E D Sverdlov |
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| Institution: | M.M. Shemyakin Institute of Bioorganic Chemistry, Moscow, USSR. |
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| Abstract: | The method of obtaining the bovine gamma-interferon gene by means of simultaneous multidirected mutagenesis of the human gamma-interferon gene is presented. The first strand of the bovine gamma-interferon gene was obtained by ligation of synthetic oligonucleotides, using the cDNA of human gamma-interferon, cloned in the single-stranded phage M13mp19 as a template. The second strand was synthesized using a large fragment of E. coli DNA-polymerase I. The double-stranded gene was then treated by restriction nucleases and cloned in a pUC-18 derived vector. The primary structure was confirmed by sequencing. |
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