Translation of chick calvarial procollagen messenger RNAs by a messenger RNA dependent reticulocyte lysate

Publisher Website
Google Scholar
Abstract
A method was developed for the extraction of RNA from chick embryo calvaria which should be generally applicable to other connective tissues. Total RNA prepared by this method was translated by a mRNA-dependent [rabbit] reticulocyte lysate into discrete pro.alpha. chains. Several criteria were used to identify these translation products, including preferential labeling with [3H]proline, appropriate migration on sodium dodecyl sulfate-polyacrylamide gels, selective sensitivity to collagenase digestion, and specific precipitability by 2 different antisera against procollagen. Data from the immunoprecipitation experiments indicated that the majority of the pro.alpha. chains contained the carboxy-terminal antigenic determinants. This translation system can be used as an assay for intact procollagen mRNAs and as a source of in vitro synthesized pro.alpha. chains for future structural analysis.