Priming reverse transcription with oligo(dT) does not yield representative samples of Mycobacterium tuberculosis cDNA

被引:12
作者
Lakey, DL
Zhang, Y
Talaat, AM
Samten, B
DesJardin, LE
Eisenach, KD
Johnston, SA
Barnes, PF [1 ]
机构
[1] Univ Texas Hlth Ctr, Ctr Pulm & Infect Dis Control, Tyler, TX 75708 USA
[2] Univ Texas Hlth Ctr, Dept Microbiol, Tyler, TX 75708 USA
[3] Univ Texas Hlth Ctr, Dept Immunol, Tyler, TX 75708 USA
[4] Univ Texas Hlth Ctr, Dept Med, Tyler, TX 75708 USA
[5] Univ Texas, SW Med Ctr, Ctr Biomed Invent, Dallas, TX 75390 USA
[6] Univ Arkansas Med Sci, Dept Pathol, Little Rock, AR 72205 USA
[7] Cent Arkansas Vet Hlth Care Syst, Little Rock, AR 72205 USA
来源
MICROBIOLOGY-SGM | 2002年 / 148卷
关键词
mycobacteria; gene expression; transcription;
D O I
10.1099/00221287-148-8-2567
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Several recent publications have suggested that oligo(dT) can prime reverse transcription of several mycobacterial mRNAs. To determine if this is the case for most Mycobacterium tuberculosis mRNA species, reverse transcription reactions of M. tuberculosis RNA were primed with oligo(dT) or with other primers that did not target polyadenylylated sequences, and the resultant cDNA product was evaluated. Priming with oligo(dT) yielded more cDNA than priming with an arbitrary primer for only 1 of 12 unrelated M. tuberculosis genes, as measured by competitive PCR. Priming with oligo(dT) yielded cDNA for only 30% of the genes primed for by 37 M. tuberculosis genome-directed oligonucleotides, as assessed by hybridization of cDNA with an M. tuberculosis microarray. These data demonstrate that priming of reverse transcription of mycobacterial mRNA with oligo(dT) does not yield representative samples of cDNA.
引用
收藏
页码:2567 / 2572
页数:6
相关论文
共 14 条
[1]   Polyadenylylation in mycobacteria: evidence for oligo(dT)-primed cDNA synthesis [J].
Adilakshmi, T ;
Ayling, PD ;
Ratledge, C .
MICROBIOLOGY-UK, 2000, 146 :633-638
[2]   Identification of differentially expressed mRNA in prokaryotic organisms by customized amplification libraries (DECAL):: The effect of isoniazid on gene expression in Mycobacterium tuberculosis [J].
Alland, D ;
Kramnik, I ;
Weisbrod, TR ;
Otsubo, L ;
Cerny, R ;
Miller, LP ;
Jacobs, WR ;
Bloom, BR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (22) :13227-13232
[3]   Fabrication of DNA microarrays using unmodified oligonucleotide probes [J].
Call, DR ;
Chandler, DP ;
Brockman, F .
BIOTECHNIQUES, 2001, 30 (02) :368-+
[4]   Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence [J].
Cole, ST ;
Brosch, R ;
Parkhill, J ;
Garnier, T ;
Churcher, C ;
Harris, D ;
Gordon, SV ;
Eiglmeier, K ;
Gas, S ;
Barry, CE ;
Tekaia, F ;
Badcock, K ;
Basham, D ;
Brown, D ;
Chillingworth, T ;
Connor, R ;
Davies, R ;
Devlin, K ;
Feltwell, T ;
Gentles, S ;
Hamlin, N ;
Holroyd, S ;
Hornby, T ;
Jagels, K ;
Krogh, A ;
McLean, J ;
Moule, S ;
Murphy, L ;
Oliver, K ;
Osborne, J ;
Quail, MA ;
Rajandream, MA ;
Rogers, J ;
Rutter, S ;
Seeger, K ;
Skelton, J ;
Squares, R ;
Squares, S ;
Sulston, JE ;
Taylor, K ;
Whitehead, S ;
Barrell, BG .
NATURE, 1998, 393 (6685) :537-+
[5]   Global burden of tuberculosis - Estimated incidence, prevalence, and mortality by country [J].
Dye, C ;
Scheele, S ;
Dolin, P ;
Pathania, V ;
Raviglione, RC .
JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION, 1999, 282 (07) :677-686
[6]   Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS) [J].
Graham, JE ;
Clark-Curtiss, JE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (20) :11554-11559
[7]   Assessment of the sensitivity and specificity of oligonucleotide (50mer) microarrays [J].
Kane, MD ;
Jatkoe, TA ;
Stumpf, CR ;
Lu, J ;
Thomas, JD ;
Madore, SJ .
NUCLEIC ACIDS RESEARCH, 2000, 28 (22) :4552-4557
[8]  
KATOCH VM, 1986, INT J LEPROSY, V54, P409
[9]   Polynucleotide phosphorylase, RNase II and RNase E play different roles in the in vivo modulation of polyadenylation in Escherichia coli [J].
Mohanty, BK ;
Kushner, SR .
MOLECULAR MICROBIOLOGY, 2000, 36 (04) :982-994
[10]   Search for genes potentially involved in Mycobacterium tuberculosis virulence by mRNA differential display [J].
Rindi, L ;
Lari, N ;
Garzelli, CA .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 258 (01) :94-101