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1994). Expression in Escherichia coli of the two subunits of the isozyme form of wheat germ protein synthesis initiation factor 4F. Purification of the subunits and formation of an enzymatically active complex. J. Biol. Chem. 269, 17454–17457. , Balasta, M. , and Goss, D. J. (1998). Wheat germ poly(A) binding protein enhances the binding affinity of eukaryotic initiation factor 4F and (iso)4F for cap analogues. Biochemistry 37, 1910–1916. Wells, D. , Tanguay, R. , and Gallie, D. R. (1998). HSP101 functions as a specific translational regulatory protein whose activity is regulated by nutrient status.

48 Daniel R. Gallie the lysate would be expected to reduce the translational advantage conferred by O. By determining whether a factor can reduce the translational advantage conferred by an RNA element, the requirement for that factor can be established. , 1998), the requirement of eIF4F or eIFiso4F could be examined using fractionated lysates. Capped-lucA50 and O-luc-A50 mRNAs were translated in PABP-dependent lysate (which was reduced in eIF4F/eIFiso4F) at an RNA concentration that provided a substantial degree of translational enhancement.

At the times indicated, 5-ml aliquots of the reaction mixture were withdrawn and analyzed by sodium dodecyl sulfate (SDS)^ 15% polyacrylamide gel electrophoresis (PAGE). An autoradiogram of the dried gel is shown. , 1998). An asterisk indicates the position of 1B. (B) Kinetics of EMCV RNA replication. EMCV RNA (10 mg/ml)programmed reaction mixtures were pulse labeled with [a-32P]CTP for1 h (the label was added 30 min before the times indicated on the figure). The arrow indicates the position of single-stranded EMCV RNA.

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Atomic Physics Revision


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