Construction of an Escherichia coli knockout strain for functional analysis of tRNA(Asp).
The specific aminoacylation of tRNA is critical for translation of the genetic code. A molecular description of aminoacylation requires knowledge of the relevant three-dimensional structures, biochemical parameters and the structure-function relationship of the synthetase and its substrate tRNA. Extensive structural and biochemical data are available on the aspartic acid system of Escherichia coli, but there is a paucity of cellular functional data. We have developed a system to overcome this deficiency by engineering an E. coli knockout tRNA(Asp) strain, thereby allowing a penetrating analysis of tRNA(Asp) structure and function under conditions that prevail in the cell.
McClain
W H
WH
Department of Bacteriology, University of Wisconsin, 1550 Linden Drive, E. B. Fred Hall, Madison, WI 53706-1567, USA. wmcclain@facstaff.wisc.edu
Gabriel
K
K
D000215:Acylation; D000926:Anticodon / Q000235:genetics; D001226:Aspartate-tRNA Ligase / Q000737:chemistry / Q000378:metabolism; D001224:Aspartic Acid / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D017353:Gene Deletion; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012349:RNA, Transfer, Asp / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
D000926:Anticodon; D012329:RNA, Bacterial; D012349:RNA, Transfer, Asp; D001224:Aspartic Acid; D001226:Aspartate-tRNA Ligase