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
- Country of Publication:
- United States
- Language:
- English
Similar Records
Closely related glycosylation patterns of recombinant human IL-2 expressed in a CHO cell line and natural IL-2
Role of binding ligand in toxic hybrid proteins: a comparison of EGF-ricin, EGF-ricin A-chain, and ricin
Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOASSAY
BIOCHEMICAL REACTION KINETICS
BODY
CARBOHYDRATES
CARBOXYLIC ACIDS
CLONING
CYSTEINE
DAYS LIVING RADIOISOTOPES
DIAGNOSTIC TECHNIQUES
DNA HYBRIDIZATION
DNA-CLONING
DRUGS
EVEN-ODD NUCLEI
GLUCOPROTEINS
GLYCOPROTEINS
HYBRIDIZATION
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
ISOTOPES
KIDNEYS
KINETICS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
METHIONINE
MONKEYS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PRIMATES
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
RADIOISOTOPES
REACTION KINETICS
SACCHARIDES
SULFUR 35
SULFUR ISOTOPES
THIOLS
TRACER TECHNIQUES
VERTEBRATES