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Volume 11, Number 2—February 2005
Research

Isolation of Waddlia malaysiensis, A Novel Intracellular Bacterium, from Fruit Bat (Eonycteris spelaea)

Paul K.B. Chua*†, John E. Corkill*, Poh Sim. Hooi‡, Soo Choon Cheng‡, Craig Winstanley*, and C. Anthony Hart*Comments to Author 
Author affiliations: *University of Liverpool, Liverpool, United Kingdom; †National Public Health Laboratory, Kuala Lumpur, Malaysia; ‡University of Malaya, Kuala Lumpur, Malaysia

Main Article

Table 1

Oligonucleotide primers for PCR and sequencing*

Gene target Primer sequence
PCR
16S rRNA (1,526 bp from ref. 20)
16S-FOR 5′ AGA GTT TGA TCC TGG 3′
16S-REV 5′ TAC CTT GTT ACG ACT T 3′
Tm = 55ºC
16S rRNA signature sequence (298 bp from ref. 21)
16S1GF 5′ CGG CGT GGA TGA GGC AT 3′
16S1GR 5′ TCA GTC CCA GTG TTG GC 3′
Tm = 51º C
16S – 23S rRNA signature sequence (1 kbp from ref. 21)
16SF2 5′ CCG CCC GTC ACA TCA TGG 3′
23S1GR 5′ TGG CTC ATC ATG CAA AAG GCA 3′
Tm = 61º C
23S rRNA signature sequence (627 bp: domain I from ref. 21)
23S1GR 5′ TGG CTC ATC ATG CAA AAG GCA 3′
Tm = 61º C
MurA signature sequence (690 bp from ref. 22)
murA-for 5′ GTN GGN GCN ACN GAR AA 3'
murA-rev 5′ GCC ATN ACR TAN GCR AAN CCN GC 3′
Tm = 55º C
sctN (331 bp)
sctN FOR 5' AGA RGG AAT GAA ACG TTC 3'
sctN REV 5' GGC TCR TTC ATA TCA TC 3'
Tm = 58º C
Sequencing
16S rRNA (1,526 bp)
Forward:
F1 (M13) 5 GTT TTC CCA GTC ACG ACG TTG TA 3′
F2 5′ GCT CAC CAA GGC TAA GAC GTC 3′ (277-298)
F3 5′ CTA GCT TTG ACC TGA CGC TGA T 3′ (752-774)
F4 5′ GAA TCT GCA ACT CGG CTC CAT G 3′ (1323-1345)
Reverse:
R1 (M13) 5′ TTG TGA GCG GAT AAC AAT TTC 3′
R2 5′ CAT CCT AAA TGC TGG CAA C 3' (392-373)
R3 5′ CAC CGC TAC ATG TGG AAT TCC 3′ (843-822)
R4 5′ GAT CCT CTC TAG CAC CAT ATC C 3′ (1358-1336)

*PCR, polymerase chain reaction; Tm, melting temperature.

Main Article

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Page updated: April 27, 2011
Page reviewed: April 27, 2011
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
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