Construction of a gene coding for a precursor of human tumor necrosis factor
V. G. Korobko, I. I. Selezneva, L. N. Shingarova, S. A. Filippov, V. N. Dobrynin
M. M. Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences of the USSR, Moscow
Abstract: The cDNA sequence for human tumor necrosis factor (hTNF) was reconstructed in vitro from genomic sequence. Using the oligonucleotide directed mutagenesis, a site for restriction endonuclease ClaI was introduced into the end of the first exon. The nucleotide sequence representing the second and third exons flanked with restriction sites ClaI and XhoI was obtained by means of chemicalenzymatic synthesis. Assembly of the total gene coding for precursor of hTNF was accomplished in pTNF33 plasmid containing semisynthetic gene for mature hTNF with appropriate restriction sites.
Russian Journal of Bioorganic Chemistry 1991, 17 (10):1365-1368