Development of a tRNA-dependent in vitro translation system.
A method is described for depleting rabbit reticulocyte lysates and wheat germ extracts of endogenous tRNAs by affinity chromatography using a matrix generated by coupling ethanolamine to epoxy-activated Sepharose 6B. Greater than 90% depletion of tRNA is achieved with the result that translation becomes in effect absolutely dependent on added tRNA. This depletion procedure should prove very useful for studying the influence of tRNA concentration, and the spectrum of the tRNA population, on recoding events such as programmed frameshifting and readthrough of termination codons.
Jackson
R J
RJ
Department of Biochemistry, University of Cambridge, United Kingdom. rjj@mole.bio.cam.ac.uk
Napthine
S
S
Brierley
I
I
D000818:Animals; D014176:Protein Biosynthesis; D012333:RNA, Messenger; D012343:RNA, Transfer / Q000302:isolation & purification; D011817:Rabbits; D012156:Reticulocytes; D014908:Triticum / Q000235:genetics
D012333:RNA, Messenger; D012343:RNA, Transfer