@article{199216,
  keywords = {Bacterial Proteins, Escherichia coli, beta-Galactosidase, Genetic Techniques, Coliphages, DNA Transposable Elements, Suppression, Genetic, Lac Operon, Defective Viruses, Genes, Synthetic},
  author = {Palva and Silhavy},
  title = {lacZ fusions to genes that specify exported proteins: a general technique},
  abstract = { We have devised a general, one-step technique for isolation of strains in which the gene coding for an exported protein is fused to the gene for beta-galactosidase (lacZ). These fusions specify a hybrid protein comprised of an NH2-terminal portion of the exported protein and a large functional COOH-terminal portion of beta-galactosidase. The fusions are constructed with a derivative of the MudII (lac, Ap) phage. To overcome the lethality that is often associated with the expression of such a hybrid gene, we have recombined an early lacZ nonsense mutation onto this phage. With the use of strains that carry a temperature-sensitive nonsense suppressor, expression of the full-length hybrid protein can be controlled by varying the growth temperature. We demonstrated the utility of this technique by isolating a series of fusions to a gene, ompA, coding for a major outer membrane protein. As expected, strains containing these fusions are not viable under conditions that permit synthesis of a functional nonsense suppressor. Accordingly, this method should also be useful for direct selection of export-defective mutants. },
  year = {1984},
  journal = {Mol Gen Genet},
  volume = {194},
  pages = {388-94},
  issn = {0026-8925},
  doi = {10.1007/BF00425549},
  language = {eng},
}