Generation of various amino acids mutants in the trpR gene of Escherichia coli by site-directed mutagenesis.
Tryptophan repressor (trpR) gene lacks various amino acid codons. To establish these codons in the trpR gene, we created the mutants by site-directed mutagenesis in the trpR gene of pHK1 plasmid. The interested regions of trpR gene were amplified, cloned in pT7-5 plasmid and transformed in to the cells harboring pGP1-2 plasmid. These plasmid products were labeled with (35)S Met, and following sequencing we observed the presence of mutants for cysteine, glycine, serine and lysine in the trpR gene of E. coli. Therefore, using these approach mutants in various genes of E. coli could be established and used as a tool to study translational bypassing in trpR gene of E. coli.
Marutha Ravindran
C R
CR
Department of Pharmacology, University of Texas Health Science Center, San Antonio, Texas 78229, USA. ravindran@uthscsa.edu
D019943:Amino Acid Substitution / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D014176:Protein Biosynthesis; D012097:Repressor Proteins / Q000235:genetics*; D029968:Escherichia coli Proteins
D001426:Bacterial Proteins; D012097:Repressor Proteins; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins