Expression of human cathepsin B protein in Escherichia coli
- 7 November 1988
- journal article
- Published by Wiley in FEBS Letters
- Vol. 239 (2) , 219-222
- https://doi.org/10.1016/0014-5793(88)80920-7
Abstract
A cDNA fragment containing the coding sequence for the mature enzyme of human lysosomal proteinase cathepsin B was inserted in the pET plasmid expression vectors, so that it was placed under the control of transcription and translation signals from bacteriophage T7. Upon induction, cathepsin B antigen was detected by in situ immunoscreening of lysed E. coli and by Western blot analysis of bacterial lysates. To our knowledge this is the first report of abundant synthesis of cloned cathepsin B in any expression system. Subfragments of cathepsin B can also be generated by this technique and will be used to study cathepsin B structure and function.Keywords
This publication has 17 references indexed in Scilit:
- Identification of latent procathepsins B and L in microsomal lumen: Characterization of enzymatic activation and proteolytic processing in vitroArchives of Biochemistry and Biophysics, 1988
- Intracellular transport and processing of lysosomal cathepsin BBiochemical and Biophysical Research Communications, 1987
- Expression of human factor IX and its subfragments in Escherichia coli and generation of antibodies to the subfragmentsBiochemistry, 1987
- T7 lysozyme inhibits transcription by T7 RNA polymeraseCell, 1987
- [50] Use of antibodies to screen cDNA expression libraries prepared in plasmid vectorsPublished by Elsevier ,1987
- Vectors for selective expression of cloned DNAs by T7 RNA polymeraseGene, 1987
- Nucleotide and predicted amino acid sequences of cloned human and mouse preprocathepsin B cDNAs.Proceedings of the National Academy of Sciences, 1986
- Reversible staining and peptide mapping of proteins transferred to nitrocellulose after separation by sodium dodecylsulfate-polyacrylamide gel electrophoresisAnalytical Biochemistry, 1986
- Amino acid sequence of human liver cathepsin BFEBS Letters, 1985
- ‘ATG vectors’ for regulated high-level expression of cloned genes in Escherichia coliGene, 1985