Silencing of genes flanking the P1 plasmid centromere

Science

Volumn 283, Issue 5401, 1999, Pages 546-549

  Rodionov, Oleg ^a   ŁObocka, Małgorzata ^a,b   Yarmolinsky, Michael ^a  

^a NATIONAL CANCER INSTITUTE   (United States)

^b INSTITUTE OF BIOCHEMISTRY AND BIOPHYSICS   (Poland)

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; CENTROMERE; DNA FLANKING REGION; GENE EXPRESSION REGULATION; GENE REPLICATION; GENE SEGREGATION; PLASMID; PRIORITY JOURNAL; PROTEIN STABILITY; REPORTER GENE; SILENCER GENE; TRANSCRIPTION REGULATION;

BACTERIA (MICROORGANISMS); UNIDENTIFIED BACTERIOPHAGE;

EID: 0033593587     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.283.5401.546     Document Type: Article

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43. Template DNAs were purified with a high pure PCR template preparation kit (Boehringer Mannheim) and, unless otherwise indicated, used at a dilution of 1:850. PCR reactions were performed with 100 pmoles of each primer and 0.25 units of Taq polymerase in B buffer (Promega) in 50-μl volumes as follows: An initial denaturation at 95°C for 2 min was followed by 25 cycles with denaturation for 30 s at 95°C, annealing for 30 s at 61°C (at 54°C with primers directed against P1 DNA), polymerization for 1 min at 72°C, and a final 2-min extension at 72°C.
44. Single-letter abbreviations for the amino acid residues are as follows: D, Asp; E, Glu; G, Gly; I, Ile; K, Lys; L, Leu; P, Pro; Q, Gln; R, Arg; T, Thr; and V. Val.
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48. We thank D. K. Chattoraj for valuable discussions, bacterial strains, plasmids, and antibody to RepA, and, with R. A. Weisberg, for critical readings of early versions of the manuscript; K. Gerdes for permission to cite unpublished material; B. E. Funnell for a sample of highly purified ParB; and D. C.-H. Lin for the cross-linking immunoprecipation protocol that he developed in the A. D. Grossman laboratory. O.R. and M.L. were supported by Fogarty postdoctoral fellowships.