Molecular cloning and expression in mammalian cells of ricin B chain
In these studies, the cDNA encoding the B chain of 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 secretion of a protein with a Mr of 30-32,000 which 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. The amount of recombinant B chain secreted by the COS-M6 cells was determined by radioimmunoassay to be 1-10 ng/ml of media. Virtually all the recombinant B chain formed active ricin when mixed with native A chain; it could also bind as effectively as native B chain to the galactose-containing glycoprotein, asialofetuin. 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:
- Texas Univ., Houston, TX (USA). Health Science Center
- OSTI ID:
- 5518248
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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GLUCOPROTEINS
DNA-CLONING
RADIOIMMUNOASSAY
ANIMAL CELLS
BIOCHEMICAL REACTION KINETICS
CYSTEINE
KIDNEYS
METHIONINE
MONKEYS
SULFUR 35
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOASSAY
BODY
CARBOHYDRATES
CARBOXYLIC ACIDS
CLONING
DAYS LIVING RADIOISOTOPES
DIAGNOSTIC TECHNIQUES
DNA HYBRIDIZATION
DRUGS
EVEN-ODD NUCLEI
GLYCOPROTEINS
HYBRIDIZATION
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
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KINETICS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
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RADIOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
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THIOLS
TRACER TECHNIQUES
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550201* - Biochemistry- Tracer Techniques