Turley et al 2000

Turley et al 2000 - Transport of Peptide-MHC Class II...

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DOI: 10.1126/science.288.5465.522 , 522 (2000); 288 Science et al. Shannon J. Turley, Developing Dendritic Cells Transport of Peptide-MHC Class II Complexes in www.sciencemag.org (this information is current as of January 15, 2008 ): The following resources related to this article are available online at http://www.sciencemag.org/cgi/content/full/288/5465/522 version of this article at: including high-resolution figures, can be found in the online Updated information and services, http://www.sciencemag.org/cgi/content/full/288/5465/522#otherarticles , 12 of which can be accessed for free: cites 26 articles This article 224 article(s) on the ISI Web of Science. cited by This article has been http://www.sciencemag.org/cgi/content/full/288/5465/522#otherarticles 79 articles hosted by HighWire Press; see: cited by This article has been http://www.sciencemag.org/cgi/collection/immunology Immunology : subject collections This article appears in the following http://www.sciencemag.org/about/permissions.dtl in whole or in part can be found at: this article permission to reproduce of this article or about obtaining reprints Information about obtaining registered trademark of AAAS. is a Science 2000 by the American Association for the Advancement of Science; all rights reserved. The title Copyright American Association for the Advancement of Science, 1200 New York Avenue NW, Washington, DC 20005. (print ISSN 0036-8075; online ISSN 1095-9203) is published weekly, except the last week in December, by the Science on January 15, 2008 www.sciencemag.org Downloaded from
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proteins appear to be paralogs that arose by a gene duplication in Eukarya. The GenBank accession numbers for the aNOP56 and aFIB protein sequenc- es from S. acidocaldarius are AF201092 and AF201093, respectively. 8. J. R. Chamberlain, Y. Lee, W. S. Lane, D. R. Engelke, Genes Dev . 12 , 1678 (1998). 9. P. Wu, J. S. Brockenbrough, A. C. Metcalfe, S. Chen, J. P. Aris, J. Biol. Chem . 273 , 16453 (1998). Brie±y, the primer AO30 (5 ± -CTCGAGATCTGGATCCGGG-3 ± ) was 5 ± phosphorylated with T4 polynucleotide kinase and adenosine triphosphate (ATP) and blocked at the 3 ± end with terminal deoxynucleotidyl transferase (Gibco BRL) and dideoxycytidine triphosphate (ddCTP). The modi²ed oligo was then ligated to gel-puri²ed sRNA for 16 hours at 4°C. The ligation products were reverse transcribed with Thermoscript RT (Gibco BRL) at 55°C for 30 min, with AO31 (5 ± -CCCGGATCCAGATCTCGAG-3 ± ) as primer. The RNA template was hydrolyzed with ribonuclease H, and the cDNA strand was extended with deoxy-ATP with terminal deoxynucleotidyl transferase. The ex- tended cDNA strand was used as template for PCR (95°C denaturation, 65°C hybridization, 72°C exten- sion, 30 cycles) with AO31 and AO32 [5 ± -GCGAAT- TCTGCAG( T) 30 -3 ± ] as primers. The DNA products were cleaved with Pst I and Xho I, ligated between the Pst I and Xho I sites of plasmid pSP72, and transformed into E. coli . Plasmids were isolated and their sequences were determined.
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