Question

Is it possible for bacterial cell components used in translation such as Ribsomes, tRNA, and aminos acids, to be used with the addition of eukaryotic mRNA to produce a protein in the PCR system? Please explain as much as you can!

Answer 1

The efficiency of translation of a eukaryotic mRNA using an in vitro translation system derived from bacteria will be less. The reason is that the ribosome binding site of bacterial mRNA is the Shine-Dalgarno sequence located 4-7 bases upstream of the initiation codon. The ribosome binding site of the eukaryotic mRNA is the methylated cap that the bacterial ribosome do not recognize. However, if the AUG codon is preceded by a small part of the poly purine Shine-Dalgarno sequence, the bacterial ribosome may initiate translation resulting the production of the protein. A eukaryotic messenger can be efficiently translated in an in vitro translation system derived from bacteria when the mRNA possesses a bacterial ribosome binding site upstream of initiation codon. Such an mRNA will be efficiently translated by bacterial ribosomes using amino acyl tRNAs, initiation, elongation and termination factors of bacteria.

PCR is not an in vitro translation system. It is used for the in vitro amplification of any piece of DNA using appropriate primers, dNTPS and a thermostable DNA polymerase.