Preparation and bacterial expression of a mutant gene for human lymphotoxin
Korobko, V.G.; Davydov, I.V.; Dobrynin, V.N.; Pustoshilova, N.M.; Lebedev, L.R.; Gileva, I.P.; Petrenko, V.A.
Bioorganicheskaia Khimiia 19(4): 414-419
1993
ISSN/ISBN: 0132-3423 PMID: 8494564 Document Number: 422308
Using the oligonucleotide directed mutagenesis, a human lymphotoxin (TNF beta) mutant gene lacking 21 N-terminal codons has been obtained. Recombinant plasmid pLT21 for expression of the mutant gene has been constructed. The mutant gene in the plasmid was placed under control of a tandem of constitutive promoters from coliphage T7. A simple procedure for isolation of recombinant protein was developed. The procedure allows to obtain the highly purified biologically active mutant protein with a good yield. During biosynthesis the recombinant protein undergoes a posttranslational processing resulted in the cleavage of N-terminal methionine and leucine residues.