title: High level synthesis of biologically active recombinant trichosanthin in Escherichia coli.
abstract: Two forms of recombinant trichosanthin (rTCS) were synthesized in high levels in Escherichia coli by putting the TCS cDNA under the control of a T7 RNA polymerase-directed promoter. Purification schemes were developed to isolate the recombinant protein from both soluble and insoluble fractions. Form I rTCS possessed the mature TCS sequence and had similar biological activities as the natural protein. Its IC50 was approximately 0.13 nM in an in vitro rabbit reticulocyte translational system and a dose of around 35 micrograms protein per 25 g body weight was sufficient to induce complete abortion in mice. Form II rTCS had a propeptide of 19 aa at the C-terminus and was five times less active than Form I in inhibiting protein synthesis by a rabbit reticulocyte lysate.
authors.affiliation: Department of Biochemistry, Chinese University of Hong Kong, Shatin, NT.
authors.forename: R H
authors.initials: RH
authors.lastname: Zhu
authors.forename: T B
authors.initials: TB
authors.lastname: Ng
authors.forename: H W
authors.initials: HW
authors.lastname: Yeung
authors.forename: P C
authors.initials: PC
authors.lastname: Shaw
mesh_terms: D000019:Abortifacient Agents / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis / Q000187:drug effects; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D012270:Ribosomes / Q000187:drug effects; D012995:Solubility; D015978:Trichosanthin / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000019:Abortifacient Agents; D011994:Recombinant Proteins; D015978:Trichosanthin