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Stability and detection of recombinant pre‐pro‐concanavalin A after cytoplasmic expression in Escherichia coli
Author(s) -
Min Wang,
Jones D.Hugh
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80265-i
Subject(s) - canavalia ensiformis , concanavalin a , escherichia coli , complementary dna , microbiology and biotechnology , terminator (solar) , recombinant dna , biology , transcription (linguistics) , cytoplasm , blot , chemistry , biochemistry , gene , in vitro , ionosphere , linguistics , physics , philosophy , astronomy
The cDNA for pre‐pro‐concanavalin A (pre‐pro‐Con A) from Canavalia ensiformis was used to construct two cytoplasmic expression vectors: pKconA (no transcription terminator) and pTKconA (containing the cry transcription terminator). The latter produced 2‐ to 3‐fold greater amounts of pre‐pro‐Con A. This product containing the plant signal can be detected by Western blotting only after electrophoretic transfer in the presence of SDS, indicating reduced solubility. The signal is not removed and pre‐pro‐Con A is clearly stable after expression in E. coli JM109. The protein is cleaved and ligated as in the plant, in contrast to a recent report.