Nucleotide sequence of the phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes Tü494 and its expression in Nicotiana tabacum.
The phosphinothricin (Pt) N-acetyltransferase gene (pat) of Streptomyces viridochromogenes Tü494 is located on a 0.8-kb BglII fragment [Strauch et al., Gene 63 (1988) 65-74]. By sequencing a 1.3-kb BglII-SstII fragment, an open reading frame representing the pat gene was found. It encodes a polypeptide of 183 amino acids with an Mr of 20,621. The base composition of the pat gene is typical for Streptomyces [70.1 mol% (G + C) in total and 93.5 mol% (G + C) in the third position]. Translation of pat is initiated by a GTG codon which was identified by frameshift mutations in Escherichia coli as well as in Streptomyces lividans. Significant homology of the pat gene was found to the bialaphos-resistance gene (bar) of Streptomyces hygroscopicus [Thompson et al., EMBO J. 9 (1987) 2519-2523]. However, variations were detected in the 5'-noncoding region of the two resistance genes which may reflect differences in regulation. Since Pt is a potent herbicide, the pat gene was modified and introduced into Nicotiana tabacum by Agrobacterium-mediated leaf-disc transformation. The GTG start codon of pat was replaced by ATG. Subsequently the modified pat-coding region was fused to the 35S promoter of the cauliflower mosaic virus. Transgenic plants could directly be selected on Pt-containing medium.
Wohlleben
W
W
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, F.R.G.
Arnold
W
W
Broer
I
I
Hillemann
D
D
Strauch
E
E
Pühler
A
A
D000123:Acetyltransferases / Q000235:genetics*; D001483:Base Sequence; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D010957:Plasmids; D015183:Restriction Mapping; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014026:Nicotiana / Q000235:genetics
D004274:DNA, Recombinant; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase