1990
Nucleic Acids Research, Vol. 18, No. 21 6435
Oxford University Press
Nucleotide brasilense
sequence
of the nodG
gene
of Azospirfflum
M.Delledonne, R.Porcari and C.Fogher Istituto di Genetica, Universita Cattolica, 29100 Piacenza, Italy Submitted September 26, 1990
EMBL accession no. X52913
A 12 kb DNA segment of the A. brasilense Sp7 genome, that shows homology with an heterologous R. meliloti nod region (1) has been cloned and partially sequenced. The construction of the physical map of the cosmid with this fragment and the DNA sequence analysis of some subclones has allowed the positioning and the identification of homology with nodG from R. meliloti 41. The nucleotide sequence of the putative nodG gene of A. brasilense shares 60% homology with the nucleotide sequence of R. meliloti nodG (2). The sequence has a ORF of 765 bp coding for a protein of 255 aa residues and 27.3 kd MW. The predicted amino acid sequence shows an overall 33 % homology with R. m. nodG and two highly conserved regions (residue 68 to 93 and 133 to 190) with, respectively, 86 and 88% similarity. The nucleotide sequence of the A. b. nodG gene was compared against the nucleotide sequence data base. One sequence, coding for the actfI gene of Streptomyces coelicolor homologous to
1 51 101 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901
known oxidoreductases, shows a 63 % homology (3). The region upstream of the ATG start codon shows homology to the 16S rRNA of E. coli, and a large inverted repeat of 43 residues is present after the stop codon. The presence of the ribosome binding site, the high G+C percent inside the ORF (65.5), and the G or C preferential presence as third base in codons (82%) give strong indications as coding frame of this DNA sequence (4). Northern blot analysis indicates that the gene is constitutively expressed in A. brasilense and that the signal is enhanced in presence of root extracts.
REFERENCES 1. Fogher,C. et al. (1985) FEMS Microbiol. Len. 30, 245 -249. 2. Debelle,F. and Sharma,S.B. (1986) Nucl. Acids Res. 14, 7453. 3. Hallar,S.E. et al. (1988) Gene 74, 305-320. 4. Gribskov,M. et al. (1983) Nucl. Acids Res. 11, 539-545.
AACCCCTTTG GCCGTTTCTA CACCGGTGCC AGGACGGTTG GCCGCCTGGC GGAAGGCGAC TCGAGGCCGA ACCCGCGACA CATCGCCACC CGAAGATGGC AACGGCGTGA CGGCGTGATC GCGTCACCGT ATGGCCATCC AGCGCCTGGG TCGGAGATCG CCTGAACTAC ATCA^ GCCCGTTC ACGGCTCG T GCCGTGTGCC TTTCGCTCTC CGTCATATTC
CAGCATCCGT GCTTGCCTCA TCTCAGAAGA CATCTGCCAG TGCCGAACTT GGCTTCAAGT CAAGGCGATG TGGTGAACAA AGGCCCAGTG CGTCACCCAG TCATCAACAT AACTACTCGG CGCCGAGCTG ACATCGGCAC CACCGACAGC GCGCGGTGTC ACCCTGAACA
AACAAGAAGC CCAGGAGCTG TCGCGCTCGT GCCCTGGCCA CGAGCCGGCC TCTACGTGGC GTCGCCAAGA CGCCGGCATC GGATGCGGTT CAGGTGTCGC CTCGTCGGTG CCGCCAAGGC GCGACCAAGG CGACATGGTC GTCCCGATGA CTACCTCGCG TCAACGGCGG
CTTCTGCGGT TTACTGGAGC ATGGGCGGTC CATCGTGGCG TGGGCCAGCA GTTTCCGACT CCTGGGTCCG AGTTCTTCGC AATCTGAGCA CGAGCGCGGC AGGGCCAGGC GGCTTCACCA CAACGCCATC GTGAGACATC CCGTCCGGAC CCGGCTACGT CAGTAAGGCA
TCACCAGCGG
G5ITCC4la
CCGGACGC
GATTGTAEI TCGGCACCGC GCGAACTGCC GGAAGCCCTC TCGAAAGCTG GTGGACATCC GAAGATGACA GCCTGTTCAA TGGGGCCGCA CGGCCAGACC AGGCGCTGGC GCCCCGGGCT CGCCAGCGAT GAGATCGGCG CACCGGCTCG CCGGAAGCAA
CTTCAAAGTA AAGCGCCC
Nucleic Acids Research, Vol. 18, No. 21 6435
Oxford University Press
Nucleotide brasilense
sequence
of the nodG
gene
of Azospirfflum
M.Delledonne, R.Porcari and C.Fogher Istituto di Genetica, Universita Cattolica, 29100 Piacenza, Italy Submitted September 26, 1990
EMBL accession no. X52913
A 12 kb DNA segment of the A. brasilense Sp7 genome, that shows homology with an heterologous R. meliloti nod region (1) has been cloned and partially sequenced. The construction of the physical map of the cosmid with this fragment and the DNA sequence analysis of some subclones has allowed the positioning and the identification of homology with nodG from R. meliloti 41. The nucleotide sequence of the putative nodG gene of A. brasilense shares 60% homology with the nucleotide sequence of R. meliloti nodG (2). The sequence has a ORF of 765 bp coding for a protein of 255 aa residues and 27.3 kd MW. The predicted amino acid sequence shows an overall 33 % homology with R. m. nodG and two highly conserved regions (residue 68 to 93 and 133 to 190) with, respectively, 86 and 88% similarity. The nucleotide sequence of the A. b. nodG gene was compared against the nucleotide sequence data base. One sequence, coding for the actfI gene of Streptomyces coelicolor homologous to
1 51 101 151 201 251 301 351 401 451 501 551 601 651 701 751 801 851 901
known oxidoreductases, shows a 63 % homology (3). The region upstream of the ATG start codon shows homology to the 16S rRNA of E. coli, and a large inverted repeat of 43 residues is present after the stop codon. The presence of the ribosome binding site, the high G+C percent inside the ORF (65.5), and the G or C preferential presence as third base in codons (82%) give strong indications as coding frame of this DNA sequence (4). Northern blot analysis indicates that the gene is constitutively expressed in A. brasilense and that the signal is enhanced in presence of root extracts.
REFERENCES 1. Fogher,C. et al. (1985) FEMS Microbiol. Len. 30, 245 -249. 2. Debelle,F. and Sharma,S.B. (1986) Nucl. Acids Res. 14, 7453. 3. Hallar,S.E. et al. (1988) Gene 74, 305-320. 4. Gribskov,M. et al. (1983) Nucl. Acids Res. 11, 539-545.
AACCCCTTTG GCCGTTTCTA CACCGGTGCC AGGACGGTTG GCCGCCTGGC GGAAGGCGAC TCGAGGCCGA ACCCGCGACA CATCGCCACC CGAAGATGGC AACGGCGTGA CGGCGTGATC GCGTCACCGT ATGGCCATCC AGCGCCTGGG TCGGAGATCG CCTGAACTAC ATCA^ GCCCGTTC ACGGCTCG T GCCGTGTGCC TTTCGCTCTC CGTCATATTC
CAGCATCCGT GCTTGCCTCA TCTCAGAAGA CATCTGCCAG TGCCGAACTT GGCTTCAAGT CAAGGCGATG TGGTGAACAA AGGCCCAGTG CGTCACCCAG TCATCAACAT AACTACTCGG CGCCGAGCTG ACATCGGCAC CACCGACAGC GCGCGGTGTC ACCCTGAACA
AACAAGAAGC CCAGGAGCTG TCGCGCTCGT GCCCTGGCCA CGAGCCGGCC TCTACGTGGC GTCGCCAAGA CGCCGGCATC GGATGCGGTT CAGGTGTCGC CTCGTCGGTG CCGCCAAGGC GCGACCAAGG CGACATGGTC GTCCCGATGA CTACCTCGCG TCAACGGCGG
CTTCTGCGGT TTACTGGAGC ATGGGCGGTC CATCGTGGCG TGGGCCAGCA GTTTCCGACT CCTGGGTCCG AGTTCTTCGC AATCTGAGCA CGAGCGCGGC AGGGCCAGGC GGCTTCACCA CAACGCCATC GTGAGACATC CCGTCCGGAC CCGGCTACGT CAGTAAGGCA
TCACCAGCGG
G5ITCC4la
CCGGACGC
GATTGTAEI TCGGCACCGC GCGAACTGCC GGAAGCCCTC TCGAAAGCTG GTGGACATCC GAAGATGACA GCCTGTTCAA TGGGGCCGCA CGGCCAGACC AGGCGCTGGC GCCCCGGGCT CGCCAGCGAT GAGATCGGCG CACCGGCTCG CCGGAAGCAA
CTTCAAAGTA AAGCGCCC
