3′-Terminal labelling of RNA with T4 RNA ligase

England, Thomas E.; Uhlenbeck, Olke C.

doi:10.1038/275560a0

Cited by 554 publications

(240 citation statements)

References 23 publications

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“…Prior to RNase H cleavage in the presence ofoligo(dT)10 (Agranovsky etal., 1982), total virion RNA was 3'-labelled by incubation with T4 RNA ligase and [0t-32p]pCp (England & Uhlenbeck, 1978). The labelled products of the RNase H cleavage were analysed in 8% thin sequencing gels and visualized by autoradiography.…”

Section: Aminoacylation Of Viral Rnas and Analysis Of Internal Poly(a)mentioning

confidence: 99%

Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome

Agranovsky

Karasev

Novikov

et al. 1992

Journal of General Virology

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RNA from the Hungarian isolate of poa semilatent virus (PSLV) directed in vitro synthesis of 120K, 75K, 25K (coat protein) and 20K polypeptides. In vitro translation of PSLV RNA was blocked by the cap analogue, m7Gpp, thus suggesting that the virus RNA was capped. PSLV RNA could be aminoacylated with [~4C]tyrosine in vitro. The sequence of 1-5 kb from the 3' end of the PSLV RNA y component revealed two open reading frames (ORFs) separated by a uridinerich intergenic region. The putative product of the incomplete 5'-proximal ORF showed a close amino acid sequence similarity with the C-terminal segment of the 7a protein (putative RNA replicase) encoded in the barley stripe mosaic virus (BSMV) RNA 7, and the 20K product of the 3'-proximal ORF was found to be related to the 17K 7b product of BSMV. The sequence of 0-8 kb from the 3' end ofPSLV RNA fl encompassed two (incomplete) overlapping ORFs whose putative products are related to the tic and fld proteins encoded in the similarly arranged ORFs of BSMV RNA ft. Nucleotide sequence homology between the respective parts of the two hordeivirus genomes was restricted to the ORF for 7a, the spacer between the ORFs for ya and yb, and the 3' non-coding region, particularly the 95 nucleotide segment at the 3' end representing a tRNAlike structure. Despite limited sequence conservation beyond this segment, the entire 3' non-coding region of PSLV RNA could be folded in a tight pseudoknotted structure closely resembling that of BSMV RNA. Surprisingly, the 'signature' sequence typical for BSMV RNA, internal polydisperse poly(A) intercalated between the coding part and the 3' tRNA-like structure, was not detected in the PSLV genome. Instead, the virus RNA contained several oligoadenylate stretches spaced by other residues, close to the junction of its coding and 3' non-coding portions.

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Section: Aminoacylation Of Viral Rnas and Analysis Of Internal Poly(a)mentioning

confidence: 99%

Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome

Agranovsky

Karasev

Novikov

et al. 1992

Journal of General Virology

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“…Genomic dsRNAs were labelled at their 3' termini using 3',5' bis-cytidine [32p]diphosphate (pCp; 3000 Ci/mnml ; Amersham) and T4 R NA ligase (Takara Shuzo) as described by England & Uhlenbeck (1978).…”

Section: E T H O D Smentioning

confidence: 99%

Viruses in the Phytoreovirus Genus of the Reoviridae Family have the Same Conserved Terminal Sequences

Kudo

Uyeda

Shikata

1991

Journal of General Virology

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“…The RNA fragments were 5 '-labeled with polynucleotide kinase and [y-32P]ATP. Nucleotide sequencing was performed according to Maxam [9] after denaturation with CH3HgOH. The labeled RNA was hybridized with the purified cDNA insert and after ethanol precipitation, the hybrids were subjected to ribonuclease T2 digestion.…”

Section: Purification Of the Viral Rnamentioning

confidence: 99%

Stable secondary structures at the 3'‐end of the genome of yellow fever virus (17 D vaccine strain)

1985

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3′-Terminal labelling of RNA with T4 RNA ligase

Cited by 554 publications

References 23 publications

Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome

Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome

Viruses in the Phytoreovirus Genus of the Reoviridae Family have the Same Conserved Terminal Sequences

Stable secondary structures at the 3'‐end of the genome of yellow fever virus (17 D vaccine strain)

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