Researchers interested in the role of Guanine Nucleotide Binding Proteins in translation expressed two of these proteins in E. Coli fused with a His-tag (GProt-1-his, and GProt-2 His); then completed electrophoretic mobility shift assays using either radiolabelled IRES RNA or hairpin subdomains of this IRES (RNA1, RNA2, and RNA3), or control RNA lacking the secondary structure of the IRES but of identical length and GC content. Their observations are reproduced below: (B) Describe in detail the results shown and indicate what can be inferred in regards to every G- protein and every RNA species assayed. (C) Design an confirmatory experiment that verifies the conclusions obtained from numeral (B), above, using fluorescent molecules or proteins. Describe in detail the fundamental principle of the experiment and the experimental design. IRES RNA RNA 1 - RNA 2 - RNA 3 - Control RNA.

1. Researchers interested in the role of Guanine Nucleotide Binding Proteins in translation expressed
two of these proteins in E. Coli fused with a His-tag (GProt-1-his, and GProt-2 His); then
completed electrophoretic mobility shift assays using either radiolabelled IRES RNA or hairpin
subdomains of this IRES (RNA1, RNA2, and RNA3), or control RNA lacking the secondary
structure of the IRES but of identical length and GC content. Their observations are reproduced
below:
(B) Describe in detail the results shown and indicate what can be inferred in regards to every G-
protein and every RNA species assayed.
(C) Design an confirmatory experiment that verifies the conclusions obtained from numeral (B),
above, using fluorescent molecules or proteins. Describe in detail the fundamental principle of the
experiment and the experimental design.
IRES RNA RNA 1 - RNA 2 - RNA 3 - Control RNA