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Molecular cloning and expression of ESAT-6 antigen of Mycobacterium bovis for differential diagnosis of bovine tuberculosis [electronic resource].

By:

Soliman, Y. A

Language: English Summary language: Arabic Description: P. 122-134Other title:

كلونة وتعبير الأنتيجين ESAT-6 من ميكروب السل البقرى للتشخيص التفاضلى للسل البقرى [Added title page title]

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Zagazig veterinary journal, 2007 v. 35 (1) [electronic resource].

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In: Zagazig Veterinary Journal 2007.v.35(1)Summary: In a search for developing new skin test reagents, the possibility of using ESAT-6 protein antigen as a candidate antigen for the diagnosis of bovine tuberculosis was investigated. Genomic DNA of M. bovis was extracted, purified and the esat-6 gene was amplified by PCR. The gene was then ligated to an expression vector, PQE. After transformation of an E. coli TOPO host strain with the PQE plasmid, the expression was induced using 10 mM of IPTG. A band (18 KDa) was seen in the SDS-PAGE analysis. The His-tagged r ESAT-6 antigen was then purified by metal affinity chromatography using Ni-NTA agarose.

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In a search for developing new skin test reagents, the possibility of using ESAT-6 protein antigen as a candidate antigen for the diagnosis of bovine tuberculosis was investigated. Genomic DNA of M. bovis was extracted, purified and the esat-6 gene was amplified by PCR. The gene was then ligated to an expression vector, PQE. After transformation of an E. coli TOPO host strain with the PQE plasmid, the expression was induced using 10 mM of IPTG. A band (18 KDa) was seen in the SDS-PAGE analysis. The His-tagged r ESAT-6 antigen was then purified by metal affinity chromatography using Ni-NTA agarose.

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