2.13 Electrophoretic mobility shift assay (EMSA)
The electrophoretic mobility shift assay was conducted using an RNA EMSA
Kit (BersinBio, Guangzhou, China) according to the manufacturer’s
instructions. Briefly, the full‐length CDS of MeCSP5 was
amplified by PCR and then cloned into the pCzn1‐His to express the
MeCSP5 protein in Escherichia coli DE3 strain. The recombinant
MeCSP5 protein was purified using the Ni affinity chromatography. EMSA
was performed in a reaction mixture containing MeCSP5 proteins and
biotin-labeled CRIR1 RNA probes. Negative control assays were
performed by adding 100-fold excesses of unlabeled RNA probes to the
reaction mixture. After electrophoresis and incubation, signals were
recorded and analyzed.