IRESite record type: plasmid_with_promoter_and_putative_IRES_translationally_characterized
The shape of the nucleic acid molecule translated: linear
The quality of the mRNA/+RNA sequence: our_best_guess
The mRNA/+RNA description:
In vivo CMV transcript produced from monocistronic plasmid pMC-env containing full-length (except very 5'-most
Adenosine) gypsy-env 5' UTR (nt from -841 to -511 of the original sequence) mRNA cloned upstream of the
beta-galactosidase.
The mRNA/+RNA sequence represented in the +DNA notation:
Credibility of mRNA sequence: end-to-end_sequence_reverse_engineered_and_should_match_experiment
The name of the promoter used to express this mRNA: CMV
Description of the plasmid (facultative for promoter-less plasmid records): Plasmid containing gypsy-env IRES (nt from -841 to nt -511 of the original sequence) and beta-galactosidase
reporter gene.
The in vivo produced transcripts are heterogeneous (due to any of promoter?/splicing?/cleavage?/breakage?): not tested
The in vivo produced heterogeneous transcripts occur due to alternative splicing: not tested
A promoter reported in cDNA corresponding to IRES sequence: no (not convincing)
Summary of possible issues when IRES cDNA is experimentally transcribed in vivo:
Summary of experiments studying integrity of the in vivo transcripts in a particular host:
Integrity (uniformity) of mRNA tested using Northern-blot: homogeneous_population_of_molecules_confirmed
Integrity (uniformity) of mRNA tested using RNase protection: not_tested
Integrity (uniformity) of mRNA tested using 5'-RACE: not_tested
Integrity (uniformity) of mRNA tested using primer extension : not_tested
Integrity (uniformity) of mRNA tested using RT-PCR: not_tested
Integrity (uniformity) of mRNA tested using real-time quantitative polymerase chain reaction (rtqPCR): not_tested
Integrity (uniformity) of mRNA tested using RNAi: not_tested
Integrity (uniformity) of mRNA tested using S1 nuclease mapping: not_tested
Cryptic promoter presence was confirmed by expression from a promoter-less plasmid: not_tested
Cryptic promoter presence was confirmed in an experimental setup involving inducible promoter: not_tested
Integrity (uniformity) of mRNA molecules or possible promoter presence expressed in vivo was tested using another method, please specify in Remarks: not_tested
The relative translation efficiency in % of this IRES: 100.000
Name of the plasmid used as the negative control. pbeta-actin-lacZ
The relative translation efficiency in % of the negative control: 14.700
The effect of 5'-cap analogs on translation: not tested
Rapamycin affects translation: not tested
Type of RNA subject to translation: endogenous_nuclear_RNA_Pol_II_transcript
Remarks:
In the absence of P2A (poliovirus protease 2A), the enzymatic activity of both pMC-Env and pbeta-actin-lacZ
reached similar levels. Upon P2A expression, a drastic decrease of beta-Gal expression from the
pbeta-actin-lacZ RNA (over fivefold decrease) was observed, whereas translation driven by the env 5' UTR was
moderately stimulated.
Translational data are derived from Fig. 7B.