Expression of rat renal gamma-glutamyltransferase cDNA in Escherichia coli.
To obtain the expression of rat kidney gamma-glutamyltransferase (GGT) cDNA in E. coli, plasmids containing the cDNA sequences coding for various parts of GGT were constructed. Transformation of E. coli cells by these hybrid vectors results in a production of unglycosylated recombinant proteins, immunologically recognized by specific antirat kidney GGT antibodies. Plasmid, expressing the complete coding sequence of GGT cDNA, allows the production of enzymatically active proteins localized in the periplasmic space, while the same sequence without the N-terminal hydrophobic region results in a production of cytoplasmic proteins. These recombinant proteins present a very basic isoelectric point (pI greater than 9). These results suggest that the presence of the N-terminal region seems to be necessary to direct the expressed proteins enzymatically active in the periplasmic space.
Angele
C
C
Centre du Médicament, URA CNRS N'597, Nancy, France.
Wellman
M
M
Thioudellet
C
C
Guellaen
G
G
Siest
G
G
D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D006031:Glycosylation; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000201:enzymology*; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D014169:Transformation, Bacterial; D005723:gamma-Glutamyltransferase / Q000096:biosynthesis / Q000235:genetics*
D011994:Recombinant Proteins; D004247:DNA; D005723:gamma-Glutamyltransferase; D004262:DNA Restriction Enzymes