A versatile and general prokaryotic expression vector, pLACT7.
We have previously reported the constitutive over-expression of the tRNA-guanine transglycosylase (TGT) from plasmid pTGT1 in Escherichia coli. To obtain a controllable expression system for TGT, we have subsequently cloned the tgt gene into pET21b. Though the overexpression of TGT is inducible in pET21b, the plasmid has a low copy number, a poor yield of single-stranded DNA and relies on an E. coli strain that produces T7 RNA polymerase for protein expression. We have combined the features of pTZ18U and pET21b and have constructed a versatile plasmid pLACT7 that has a high copy number, a high yield of single-stranded DNA and both the T7 and lac promoters for protein expression in a wide variety of E. coli strains.
Chong
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S
College of Pharmacy, University of Michigan, Ann Arbor 48109-1065.
Garcia
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GA
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D010430:Pentosyltransferases / Q000235:genetics*; D010957:Plasmids
D010430:Pentosyltransferases; C020837:queuine tRNA-ribosyltransferase