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 [35S]methionine and [35S]cysteine and demonstrating the secretion of a protein with a Mr of 30,000-32,000 th...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-08, Vol.84 (16), p.5640-5644
Main Authors: Chang, M.S, Russell, D.W, Uhr, J.W, Vitetta, E.S
Format: Article
Language:English
Subjects:
ADN
Amino acids
Animals
Antibodies
Biological and medical sciences
Biotechnology
Cell Line
Cell lines
Cellular immunity
CLONACION
CLONAGE
CLONING
Cloning, Molecular
Complementary DNA
Cysteine - metabolism
DNA
Fundamental and applied biological sciences. Psychology
Galactose
Gels
GENE
Gene Expression Regulation
GENES
Genetic engineering
Genetic technics
Genetic vectors
Glycoproteins - metabolism
Haplorhini
LECTINA
LECTINE
LECTINS
Methionine - metabolism
Methods. Procedures. Technologies
Molecular chains
Molecular cloning
Molecular Weight
Recombinant Proteins - genetics
Ricin - genetics
RICINUS COMMUNIS
TOXINAS
TOXINE
TOXINS
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 [35S]methionine and [35S]cysteine and demonstrating the secretion of a protein with a Mr 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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.16.5640