The nucleic acid data:
IRESite Id: 350 Version: 3
Originaly submitted by: Václav Vopálenský
Reviewed by: Martin Mokrejš Last change: 2008-06-04 15:43:36
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:
  end-to-end_full-length_mRNA
The mRNA/+RNA description: 
In vitro T3 run-off transcript containing 5' region of PV (Mahoney strain) containing the IRES (-660 to -1)
plus the region encoding the first 37 amino acids of the viral nucleocapsid cloned upstream of the luciferase
reporter RNA lacking a poly(A) tail.
The mRNA/+RNA sequence represented in the +DNA notation:


Credibility of mRNA sequence:
  end-to-end_sequence_completely_same_as_in_the_experiment
The name of the plasmid:
pPV.IRES-luc
The name of the promoter used to express this mRNA:
  T3
Description of the plasmid (facultative for promoter-less plasmid records):
Plasmid containing the major part of PV 5' UTR (-660 to -1) plus the region encoding the first 37 amino acids of the viral polyprotein (+1 to +111) and firefly luciferase coding region (luc version of the luciferase gene) without the poly(A)tract.
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:
  not tested
The abbreviated name of the donor gene or virus from which this IRES was excised and inserted into the plasmid:
PV
The origin of IRES in the plasmid:
  viral
The donor organism of the IRES segment:
Human poliovirus 1 Mahoney
The DNA sequence of the plasmid in (+) orientation annotated by its secondary structure:


Plasmid sequence verified (partially/completely) by IRESite (more details in Remarks):
  plasmid sequence confirmed by IRESite curators by restriction analysis + parts by PCR + sequencing
GenBank formatted file with annotated plasmid sequence hyperlinked from vector image map:
pPV.IRES-luc.jpg
The total number of notable open-reading frames (ORFs):
  1
Notable Open-Reading Frames (ORFs; protein coding regions) in the mRNA/+RNA sequence:
ORF
ORF position:   1
Version: 1 Last change: 2008-06-04 11:50:07
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The abbreviated name of this ORF/gene:
FFluc
The description of the protein encoded in this ORF:
firefly luciferase
The translational frameshift (ribosome slippage) involved:
  0
The ribosome read-through involved:
  no
The alternative forms of this protein occur by the alternative initiation of translation:
  not tested
The ORF absolute position (the base range includes START and STOP codons or their equivalents):
  811-2463
Remarks:
IRESite notes about verification of plasmid sequence:
The complete sequence of IRES element was determined by sequencing using M13_reverse (CAGGAAACAGCTATGAC) and
Fluc_seq_reverse (AGGAACCAGGGCGTATCTC) primers.
Citations:
Bergamini G., Preiss T., Hentze M. W. (2000) Picornavirus IRESes and the poly(A) tail jointly promote cap-independent translation in a mammalian cell-free system. RNA. 6(12):1781-1790
IRESs:
IRES:
Version: 2 Last change: 2008-06-04 11:50:07
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The IRES name:
  PV
The functional status of IRES:
  functional
The IRES absolute position (the range includes START and STOP codons or their equivalents):
  34-804
How IRES boundaries were determined:
experimentally_determined
The sequence of IRES region aligned to its secondary structure (if available):


Remarks:
DNA fragment (-660 to +111) containing the PV IRES and 37 amino acids of the capsid protein from Human
poliovirus 1 Mahoney.
Citations:
Bergamini G., Preiss T., Hentze M. W. (2000) Picornavirus IRESes and the poly(A) tail jointly promote cap-independent translation in a mammalian cell-free system. RNA. 6(12):1781-1790
The translation experiments:
Translation results:
IRESite Translation Id: 421
Version: 1 Last change: 2008-06-04 11:50:08
Originaly submitted by: Václav Vopálenský Reviewed by: Martin Mokrejš
The translation method used to study IRES function:
in vitro
The in vitro translation system:
HeLa cell lysate
The organism used for translation:
The temperature (in degrees of Celsia):
37
The relative translation efficiency in % of this IRES:
  100.000
Name of the plasmid used as the negative control.
pT3Luc
IRESite Id of the plasmid used as negative control.
  346
The relative translation efficiency in % of the negative control:
  0.800
The effect of 5'-cap analogs on translation:
no
Rapamycin affects translation:
not tested
Type of RNA subject to translation:
  exogenous_RNA_with_ApppG_cap_without_polyA_tail
Remarks:
Translation of PV.IRES RNA is unaffected by increasing amounts of m7GpppG competitor.
The translation of PV.IRES RNA is about 10-fold less efficient than the translation of the polyadenylated
counterpart PV.IRES-pA RNA.
Translational data are derived from Fig. 4A.
Citations:
Bergamini G., Preiss T., Hentze M. W. (2000) Picornavirus IRESes and the poly(A) tail jointly promote cap-independent translation in a mammalian cell-free system. RNA. 6(12):1781-1790
Last change to the database: 2015-04-16 16:45:23 GMT+1