PMID:7685296

Citation	Borel, F, Härtlein, M and Leberman, R (1993) In vivo overexpression and purification of Escherichia coli tRNA(ser). FEBS Lett. 324:162-6
Abstract	DNA fragments corresponding to the sequences of Escherichia coli tRNA(2ser) and amber suppressor tRNA(ser), were synthesized from overlapping oligonucleotides. These were interposed between a strong promotor and a synthetic transcriptional terminator to ensure the production of a transcript of the correct size. The genes of promotor, fragment and terminator were cloned into a conditional runaway replication plasmid. At temperatures below 37 degrees C this vector has a low copy number but, following a temperature shift to 42 degrees C, the copy number is no longer regulated. Using these constructs an overexpression of tRNA(ser) of about 20 times the level of the wild-type pool could be obtained (corresponding e.g. to 200 times the expression tRNA(2ser)). From these systems 10 mg quantities of tRNA(ser)s could be isolated with a serine acceptance of 1,100 pmol/A280 unit.
Links	PubMed
Keywords	Base Sequence; Escherichia coli/genetics; Genes, Bacterial/genetics; Molecular Sequence Data; Plasmids/genetics; RNA; RNA, Transfer, Ser/biosynthesis; RNA, Transfer, Ser/genetics; RNA, Transfer, Ser/isolation & purification
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