Development of an In Vivo Assay for Mistranslation-Inducing Activity of Pollutants and Characterization of Amino Acid Substitutions.
Abstract
In experiments dilrected toward developing a simple, in vivo assay for mistranslation, we have attempted to purify monoclonal antibody to 0.3 protein from large volumes of spent hybridoma. Purification was attempted by several means but no method resulted in high yields of pure antibody in radioimmune precipitation (RIP)-SDS-PAGE experiments for 0.3 protein. It was found that proteins in addition to 0.3 protein were precipitated with monoclonal antibody from ascities as well as with rabbit antibody. No contaminating, additional proteins were precipitated with monoclonal antibody from culture fluid but wer have not been able to purify and concentrate it sufficiently to be practicable in RIP assays. In experiments directed twoard determining the influence of cellular environment on mistranslation we have show that a protein synthesized in E. coli containing the plasmid pAR324 (which contains the 4.3 gene) is of the same M. W. as 0.3 protein and is precipitated by rabbit antibody to 0.3 pritein produced in this and other pAR324 - containing bacterial strains, and in B. subtilis infected with phage SPP1-0.3
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 1984
- Accession Number
- ADA154616
Entities
People
- J. B. Rice
- J. N. Reeve
Organizations
- Ohio State University