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Identification of Mycobacterium tuberculosis by Dot Blot Hybridization Using Digoxigenin-Iabeled Probe

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dc.contributor.authorKim, Eui Chong-
dc.date.accessioned2009-08-06T04:50:04Z-
dc.date.available2009-08-06T04:50:04Z-
dc.date.issued1992-09-
dc.identifier.citationSeoul J Med, Vol.33 No.3, pp. 213-218-
dc.identifier.issn0582-6802-
dc.identifier.urihttp://hdl.handle.net/10371/6239-
dc.description.abstractMycobacterium tuberculosis H37Rv ATCC 27294, restricted with EcoRI,
was used to construct a genomic library in a bacteriophage expression vector, Lambda
9111. One 1.5-kb DNA fragment was selected by picoBlue immunoscreening kit on the
basis of the ability to encode M. bovis-specific protein. Recombinant pBluescript with
the 1.5-kb DNA was constructed into EcoRI site. Denatured DNA solution was blotted
with BIO-DOT microfiltration apparatus(BIO-RAD) onto nylon membrane. The 1.5-kb
probe, labeled with digoxigenin, reacted strongly with M. tuberculosis H37Rv, M. bovis,
and 5 clinical isolates of M. tuberculosis. No hybridizations were observed with M. tuberculosis
H37Ra, M. scrofulaceum, M. intracellulare, S. au reu s, S. epidennidis, S.
marcescens, K. pneumoniae, E. coli, E. cloacae, P. vulgaris, and P. aeruginosa. When
purified DNA from M. tuberculosis H37Rv was spotted onto nylon membrane, the
1.5-kb probe was able to detect 100 ng of DNA, which corresponds to 2 X 107
mycobacteria. This dot blot hybridization method using digoxigenin-Iabeled 1.5-kb
probe may be applicable for identifying M. tuberculosis in hospital laboratories.
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dc.language.isoen-
dc.publisherSeoul National University College of Medicine-
dc.subjectMycobacterium tuberculosis-
dc.subjectDot blot hybridization-
dc.subjectDigoxigenin-
dc.subjectLambda gt 11-
dc.subjectExpression library-
dc.titleIdentification of Mycobacterium tuberculosis by Dot Blot Hybridization Using Digoxigenin-Iabeled Probe-
dc.typeSNU Journal-
dc.contributor.AlternativeAuthor김의종-
dc.citation.journaltitle서울 의대 잡지-
dc.citation.journaltitle서울 의대 학술지-
dc.citation.journaltitleSeoul Journal of Medicine-
dc.citation.endpage218-
dc.citation.number3-
dc.citation.pages213-218-
dc.citation.startpage213-
dc.citation.volume33-
Appears in Collections:
College of Medicine/School of Medicine (의과대학/대학원)Dept. of Medicine (의학과)The Seoul Journal of MedicineThe Seoul Journal of Medicine Vol. 33 No.3 (1992)
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