Cloning and expression of recombinant, functional ricin B chain
The cDNA encoding the B chain of the plant toxin ricin has been cloned and expressed in monkey kidney COS-M6 cells. The recombinant B chain was detected by labeling the transfected cells with (/sup 35/S)methionine and (/sup 35/S)-cysteine and demonstrating the secretion of a protein with a M/sub r/ of 30,000-32,000 that was not present in the medium of mock-transfected COS-M6 cells. This protein was specifically immunoprecipitated by an anti-ricin or anti-B-chain antibody and the amount of recombinant B chain secreted by the COS-M6 cells was determined by a radioimmunoassay. Virtually all of the recombinant B chain formed active ricin when mixed with native A chain; it could also bind to the galactose-containing glycoprotein asialofetuin as effectively as native B chain.These results indicate that the vast majority of recombinant B chains secreted into the medium of the COS-M6 cells retain biological function
- Research Organization:
- Univ. of Texas Health Science Center, Dallas (USA)
- OSTI ID:
- 5298007
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:16
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PROTEINS
SECRETION
RECOMBINANT DNA
DNA-CLONING
TOXINS
AUTORADIOGRAPHY
RADIOIMMUNOASSAY
ANIMAL CELLS
CYSTEINE
ELECTROPHORESIS
IODINE 125
KIDNEYS
LECTINS
METHIONINE
MOLECULAR WEIGHT
MONKEYS
SULFUR 35
AMINO ACIDS
ANIMALS
ANTIGENS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
EVEN-ODD NUCLEI
IMMUNOASSAY
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MATERIALS
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PRIMATES
RADIOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
SULFUR ISOTOPES
THIOLS
TOXIC MATERIALS
TRACER TECHNIQUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques