TABLE OF CONTENTS The central dogma of modern biology is the conversion of the genetic message in DNA to a functional mRNA (transcription) and subsequent conversion of the copied genotype to a phenotype in the form of proteins.
The process of conversion of mRNA to a functional protein is known as translation. It involves the attachment of a messenger RNA to the smaller subunit of a ribosome, the addition of the larger subunit, plus initiation by a host of other factors. The entire process can be accomplished in the absence of a cell, if all of the necessary factors are present. Unfortunately, studies on translation and post-translational changes in protein structure are rather complex. They require a heavy investment of time and equipment. To some extent, the electrophoretic identification of proteins is part of this process, and the appearance of specific proteins can be monitored during any of the developmental processes.
To study the process of translation in any meaningful way requires that reasonably purified sources of mRNA, ribosomes, and amino acids be available. In addition, there is a requirement for various factors responsible for peptide chain initiation on the ribosome.
It is definitely an advanced technique, and requires mastery of many of the techniques presented in previous chapters. It must be performed on an independent basis, since the extensive time commitment does not lend itself to typical laboratory periods.
Suspension culture of fibroblast cells (1 liter)
