[Chemical-enzymatic synthesis and cloning of DNA, coding the signal for secretion of proteins in gram-negative bacteria].
Chemical-enzymatic synthesis of an artificial gene encoding leader peptide and 22 N-terminal amino acids of mature carboxypeptidase G2 from Pseudomonas sp. followed by enterokinase signal sequence (Asp4Lys) has been accomplished. The resulted DNA was fused with semi-synthetic gene coding for polypeptide 4-157 of mature human tumour necrosis factor (TNF) and then placed under control of early promoters of T7 bacteriophage. The expression products of the construct obtained was analysed using anti-TNF anti-serum. In E.coli leader peptide was cleaved off during translocation through inner membrane and the resultant product was found in membrane fraction.
Dobrynin
V N
VN
Kalinichenko
S V
SV
Farkhutdinov
M R
MR
Filippov
S A
SA
Korobko
V G
VG
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005813:Genes, Synthetic; D006090:Gram-Negative Bacteria / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D014409:Tumor Necrosis Factor-alpha; D004247:DNA
Khimiko-fermantativnyĭ sintez i klonirovanie DNK, kodiruiushcheĭ signal dlia sekretsii belkov v gramootritsatel'nykh bakteriaiakh.