Optimal conditions for cell-free synthesis of elastin

Optimal conditions for the translation of elastin mRNA1 in a mRNA-dependent rabbit reticulocyte lysate were determined. Using total RNA isolated from embryonic chick aortae as the source of exogenous RNA, the concentrations of various components present in the translation assay were varied and the effect on elastin synthesis quantitated by immunoprecipitation. Components examined included: magnesium acetate, potassium chloride, spermidine, creatine phosphate, ATP, and GTP. In addition, it was found that heating of the RNA prior to translation significantly enhanced total protein synthesis, elastin synthesis, and the synthesis of proteins possessing molecular weights of greater than 80,000.