TY - JOUR
T1 - Determination of the nucleotide sequence for the glutamate synthase structural genes of Escherichia coli K-12
AU - Oliver, Guillermo
AU - Gosset, Guillermo
AU - Sanchez-Pescador, Ray
AU - Lozoya, Edmundo
AU - Ku, Lailig M.
AU - Flores, Noemi
AU - Becerril, Baltazar
AU - Valle, Fernando
AU - Bolivar, Francisco
N1 - Funding Information:
We wish to thank Gilbert0 Mosqueda for his help in the purification of the GOGAT enzyme. The protein sequence determination was performed by Guillermo Ramirez in the Protein/DNA Sequence Synthesis Facility of the University of Wisconsin Biotechnology Center, Madison, WI (U.S.A.), supported by funds from the Public Health Service, National Institutes of Health (Shared Equipment grant SlO-RRi684, National Cancer Institute continued support grant CA-07175 and the General Research Support grant to the University of Wisconsin Medical School) and from the University of Wisconsin Graduate School. We wish to thank Dr. Edmund0 Calva who first developed some of the purification manipulations of the GOGAT enzyme at the University of Wisconsin Biotechnology Center. G. Gosset is a recipient of a fellowship from Consejo National de Ciencia y Tecnologia, Mexico. This work was supported in part by Consejo National de Ciencia y Tecnologia, Mexico, Donativo PCCBNAL-022584.
PY - 1987
Y1 - 1987
N2 - We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.
AB - We have determined the complete nucleotide sequence of a 6.3-kb chromosomal HpaI-EcoRI fragment, that contains the structural genes for both the large and small subunits of the Escherichia coli K-12 glutamate synthase (GOGAT) enzyme, as well as the 5 ' - and 3 ' -flanking and intercistronic DNA regions. The Mrs of the two subunits, as deduced from the nucleotide (nt) sequence, were estimated as 166 208 and 52 246. Partial amino acid sequence of the GOGAT enzyme revealed that the large subunit starts with a cysteine residue that is probably generated by a proteolytic cleavage. Northern blotting experiments revealed a transcript of approximately 7300 nt, that at least contains the cistrons for both subunits. A transcriptional start point and a functional promoter were identified in the 5' DNA flanking region of the large subunit gene. The messenger RNA nontranslated leader region has 120 nt and shares identity with the leader regions of E. coli ribosomal operons, in particular around the so-called boxA sequence implicated in antitermination. Other possible regulatory sequences are described.
KW - Recombinant DNA
KW - flavoprotein
KW - functional promoter
KW - nitrogen metabolism
KW - oligodeoxynucleotide primer
KW - protein amino terminus
KW - transcriptional start point
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U2 - 10.1016/0378-1119(87)90207-1
DO - 10.1016/0378-1119(87)90207-1
M3 - Article
C2 - 3326786
AN - SCOPUS:0023551349
VL - 60
SP - 1
EP - 11
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1
ER -