[Preparation and bacterial expression of a mutant gene for human lymphotoxin].
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.
Korobko
V G
VG
Davydov
I V
IV
Dobrynin
V N
VN
Pustoshilova
N M
NM
Lebedev
L R
LR
Gileva
I P
IP
Petrenko
V A
VA
D000595:Amino Acid Sequence; D001483:Base Sequence; D003062:Codon; D004279:DNA, Viral; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D008233:Lymphotoxin-alpha / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational
D003062:Codon; D004279:DNA, Viral; D008233:Lymphotoxin-alpha
Poluchenie i bakteral'naia ékspressiia mutantnogo gena limfotoksina cheloveka.