Abstract:
:Two analog uridine triphosphates tethering additional functionality, one a primary amino group and the second a mercapto group, were prepared and tested for their compatibility with in vitro RNA selection procedures. 5-(3-Aminopropyl)uridine triphosphate (UNH(2)) as a uridine substitute was a more effective substrate for T7 RNA polymerase than 5-(2-mercaptoethyl)uridine triphosphate (USH). However, both functioned in transcription assays of 100 nt templates to generate RNA transcripts in quantities sufficient to initiate RNA selection procedures. Transcription of RNA pools with T7 RNA polymerase and UNH(2) or USH occurred with efficiencies of 43 and 29%, respectively, of the values obtained for native UTP transcription. In addition, the transcribed RNA containing roughly 25% UNH(2) residues exhibited better substrate properties for SuperScript(TM) II RNase H reverse transcriptase than did RNA transcripts containing approximately 25% of the USH analog. With either analog, both transcription and reverse transcription proceeded with high fidelity for insertion of the analog residue.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Vaish NK,Fraley AW,Szostak JW,McLaughlin LWdoi
10.1093/nar/28.17.3316keywords:
subject
Has Abstractpub_date
2000-09-01 00:00:00pages
3316-22issue
17eissn
0305-1048issn
1362-4962journal_volume
28pub_type
杂志文章abstract::In the past year, DDBJ (http://www.ddbj.nig.ac.jp) collected and released 1,956,826 entries or 1,741,313,111 bases. The released data include approximately 90,000 ESTs and cDNAs of Macaca fascicularis, and 280 million bases of mouse GSS. In addition to the data collection, we have indexed the submitted data to the Int...
journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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abstract::Although a vast amount of life sciences data is generated in the form of images, most scientists still store images on extremely diverse and often incompatible storage media, without any type of metadata structure, and thus with no standard facility with which to conduct searches or analyses. Here we present a solutio...
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更新日期:2011-04-01 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
pub_type: 杂志文章
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更新日期:2015-02-27 00:00:00
abstract::One surprisingly common element of RNA secondary structure consists of a hairpin capped by a four-base loop (or the tetraloop). Recently the 3-D structures of two RNA-tetraloops have been determined by NMR-studies. Both structures have a similar architecture: the first and the last bases of the loop form a hydrogen bo...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.19.4556
更新日期:1993-09-25 00:00:00
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journal_title:Nucleic acids research
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doi:10.1093/nar/gkx920
更新日期:2018-01-04 00:00:00
abstract::Global run-on sequencing (GRO-seq) is a recent addition to the series of high-throughput sequencing methods that enables new insights into transcriptional dynamics within a cell. However, GRO-sequencing presents new algorithmic challenges, as existing analysis platforms for ChIP-seq and RNA-seq do not address the uniq...
journal_title:Nucleic acids research
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更新日期:2014-02-01 00:00:00
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journal_title:Nucleic acids research
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更新日期:1974-08-01 00:00:00
abstract::The structure and organisation of the human N-ras gene has been determined by analysing cDNA clones derived from the two main mRNA transcripts. One clone in particular is 4.1 Kb long and originates from the larger (4.3 Kb) message. Sequence analysis of this clone has revealed that the N-ras gene consists of seven exon...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/13.14.5255
更新日期:1985-07-25 00:00:00
abstract::Minichromosome maintenance (MCM) proteins facilitate replication by licensing origins and unwinding the DNA double strand. Interestingly, the number of MCM hexamers greatly exceeds the number of firing origins suggesting additional roles of MCMs. Here we show a hitherto unanticipated function of MCM2 in cilia formatio...
journal_title:Nucleic acids research
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更新日期:2019-01-10 00:00:00
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journal_title:Nucleic acids research
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journal_title:Nucleic acids research
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abstract::The dissection of RNA editing mechanisms in PHYSARUM: mitochondria has been hindered by the absence of a soluble in vitro system. Based on our studies in isolated mitochondria, insertion of non-encoded nucleotides into PHYSARUM: mitochondrial RNAs is closely linked to transcription. Here we have fractionated mitochond...
journal_title:Nucleic acids research
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更新日期:2000-10-01 00:00:00
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更新日期:1997-06-01 00:00:00
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