Abstract
We have tested conditions for the labeling and tailing the 3'-end of RNAs with yeast poly(A) polymerase. Conditions were optimized for addition of NTP, dNTP, or ddNTP nucleotides to RNA. ATP, GTP, and UTP were useful for adding homopolymer tracts of various lengths. The nonradioactive nucleotides biotin-N6-ATP and digoxigenin-11-UTP also were used efficiently.
Full Text
The Full Text of this article is available as a PDF (501.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Amara R. R., Vijaya S. Specific polyadenylation and purification of total messenger RNA from Escherichia coli. Nucleic Acids Res. 1997 Sep 1;25(17):3465–3470. doi: 10.1093/nar/25.17.3465. [DOI] [PMC free article] [PubMed] [Google Scholar]
- England T. E., Uhlenbeck O. C. 3'-terminal labelling of RNA with T4 RNA ligase. Nature. 1978 Oct 12;275(5680):560–561. doi: 10.1038/275560a0. [DOI] [PubMed] [Google Scholar]
- Frendewey D., Keller W. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences. Cell. 1985 Aug;42(1):355–367. doi: 10.1016/s0092-8674(85)80131-8. [DOI] [PubMed] [Google Scholar]
- Hertel K. J., Herschlag D., Uhlenbeck O. C. Specificity of hammerhead ribozyme cleavage. EMBO J. 1996 Jul 15;15(14):3751–3757. [PMC free article] [PubMed] [Google Scholar]
- Huang Z., Szostak J. W. A simple method for 3'-labeling of RNA. Nucleic Acids Res. 1996 Nov 1;24(21):4360–4361. doi: 10.1093/nar/24.21.4360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kessler M. M., Henry M. F., Shen E., Zhao J., Gross S., Silver P. A., Moore C. L. Hrp1, a sequence-specific RNA-binding protein that shuttles between the nucleus and the cytoplasm, is required for mRNA 3'-end formation in yeast. Genes Dev. 1997 Oct 1;11(19):2545–2556. doi: 10.1101/gad.11.19.2545. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langer-Safer P. R., Levine M., Ward D. C. Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4381–4385. doi: 10.1073/pnas.79.14.4381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lingner J., Hendrick L. L., Cech T. R. Telomerase RNAs of different ciliates have a common secondary structure and a permuted template. Genes Dev. 1994 Aug 15;8(16):1984–1998. doi: 10.1101/gad.8.16.1984. [DOI] [PubMed] [Google Scholar]
- Lingner J., Keller W. 3'-end labeling of RNA with recombinant yeast poly(A) polymerase. Nucleic Acids Res. 1993 Jun 25;21(12):2917–2920. doi: 10.1093/nar/21.12.2917. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lingner J., Kellermann J., Keller W. Cloning and expression of the essential gene for poly(A) polymerase from S. cerevisiae. Nature. 1991 Dec 12;354(6353):496–498. doi: 10.1038/354496a0. [DOI] [PubMed] [Google Scholar]
- Lingner J., Radtke I., Wahle E., Keller W. Purification and characterization of poly(A) polymerase from Saccharomyces cerevisiae. J Biol Chem. 1991 May 15;266(14):8741–8746. [PubMed] [Google Scholar]
- Martin G., Keller W. Mutational analysis of mammalian poly(A) polymerase identifies a region for primer binding and catalytic domain, homologous to the family X polymerases, and to other nucleotidyltransferases. EMBO J. 1996 May 15;15(10):2593–2603. [PMC free article] [PubMed] [Google Scholar]
- Rosemeyer V., Laubrock A., Seibl R. Nonradioactive 3'-end-labeling of RNA molecules of different lengths by terminal deoxynucleotidyltransferase. Anal Biochem. 1995 Jan 1;224(1):446–449. doi: 10.1006/abio.1995.1068. [DOI] [PubMed] [Google Scholar]
- Schnorf M., Neuhaus-Url G., Galli A., Iida S., Potrykus I., Neuhaus G. An improved approach for transformation of plant cells by microinjection: molecular and genetic analysis. Transgenic Res. 1991 Dec;1(1):23–30. doi: 10.1007/BF02512993. [DOI] [PubMed] [Google Scholar]
- Shepard A. R., Rae J. L. Magnetic bead capture of cDNAs from double-stranded plasmid cDNA libraries. Nucleic Acids Res. 1997 Aug 1;25(15):3183–3185. doi: 10.1093/nar/25.15.3183. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tautz D., Pfeifle C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma. 1989 Aug;98(2):81–85. doi: 10.1007/BF00291041. [DOI] [PubMed] [Google Scholar]
- Thomson J. G., Gershon P. D. Use of vaccinia virus poly(A) polymerase for RNA 3'-end labeling with a chain-terminating nucleotide or a short 3' homopolymer tract. Biotechniques. 1995 Sep;19(3):416-20, 422-5. [PubMed] [Google Scholar]
- Wahle E., Martin G., Schiltz E., Keller W. Isolation and expression of cDNA clones encoding mammalian poly(A) polymerase. EMBO J. 1991 Dec;10(13):4251–4257. doi: 10.1002/j.1460-2075.1991.tb05003.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wahle E. Purification and characterization of a mammalian polyadenylate polymerase involved in the 3' end processing of messenger RNA precursors. J Biol Chem. 1991 Feb 15;266(5):3131–3139. [PubMed] [Google Scholar]
- Winter G., Brownlee G. G. 3'End labelling of RNA with 32P suitable for rapid gel sequencing. Nucleic Acids Res. 1978 Sep;5(9):3129–3139. doi: 10.1093/nar/5.9.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zaug A. J., Linger J., Cech T. R. Method for determining RNA 3' ends and application to human telomerase RNA. Nucleic Acids Res. 1996 Feb 1;24(3):532–533. doi: 10.1093/nar/24.3.532. [DOI] [PMC free article] [PubMed] [Google Scholar]