Effective method for obtaining long nucleotide chains on partially complementary templates. Processed bovine gamma-interferon gene obtained from human gamma-interferon gene

FEBS Lett. 1989 Jun 5;249(2):379-82. doi: 10.1016/0014-5793(89)80663-5.

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.

MeSH terms

  • Animals
  • Base Sequence
  • Cattle
  • Electrophoresis, Agar Gel
  • Humans
  • Interferon-gamma / genetics*
  • Molecular Sequence Data
  • Oligonucleotides / chemical synthesis*
  • Sequence Homology, Nucleic Acid
  • Templates, Genetic

Substances

  • Oligonucleotides
  • Interferon-gamma