[Study of the biogenesis and secretion of alkaline phosphatase and its mutant forms in Escherichia coli. I. Introduction of directed mutations into the alkaline phosphatase gene].
Various mutations in E. coli alkaline phosphatase gene were obtained by oligonucleotide-directed mutagenesis. They result in amino acid substitutions in the signal peptide cleavage site [Val for Ala(-1)] and in the N terminus of mature polypeptide chain: Ala for Arg(+1) and Gln for Glu(+4); Gln for Glu(+4). Enzyme activity was observed in all E. coli strains transformed by plasmids with cloned mutant genes. In addition, an amber mutation was introduced into the Arg(+1) position, and the synthesis of mutant alkaline phosphatase was shown in E. coli strains containing suppressor tRNAs specific for Ser, Gln, Tyr, Leu, Ala, Glu, Phe, Gly, His, Pro, and Cys.
Karamyshev
A L
AL
Shliapnikov
M G
MG
Khmel'nitskiĭ
M I
MI
Nesmeianova
M A
MA
Ksenzenko
V H
VH
D000469:Alkaline Phosphatase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D012343:RNA, Transfer / Q000378:metabolism
D009838:Oligodeoxyribonucleotides; D012343:RNA, Transfer; D000469:Alkaline Phosphatase
Izuchenie biogeneza i sekretsii shchelochnoĭ fosfatazy i ee mutantnykhform u Excherichia coli. I. Vvedenie napravlennykh mutatsiĭ v gen shchelochnoĭ fosfatazy.