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Immunological screening of a genomic M. bovis BCG library expressed in M. smegmatis and identification of the M. bovis BCG analog of ClpB
This work describes the screening of a M. bovis BCG cosmid library in M. smegmatis with a hyperimmune rabbit anti-BCG serum. Cross-reactive antibodies interfere with the detection of BCG specific antigens in M. smegmatis culture filtrates. We, therefore, screened parallel western blots with serum ad...
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| Published in: | Tubercle and lung disease 1998-01, Vol.78 (2), p.133-141 |
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| container_end_page | 141 |
| container_issue | 2 |
| container_start_page | 133 |
| container_title | Tubercle and lung disease |
| container_volume | 78 |
| creator | Bona, M. Nayak, R. Wu, M. Mincek, M. Ellner, J.J. |
| description | This work describes the screening of a
M. bovis BCG cosmid library in
M. smegmatis with a hyperimmune rabbit anti-BCG serum.
Cross-reactive antibodies interfere with the detection of BCG specific antigens in
M. smegmatis culture filtrates. We, therefore, screened parallel western blots with serum adsorbed with a
M. smegmatis cell lysate and unadsorbed serum. Comparison of the western blots allowed distinction between BCG specific and cross-reactive
M. smegmatis antigens. Thirty-one cosmids expressed BCG specific antigens. One of them, a hitherto undescribed 100 kDa antigen was subcloned, sequenced and expressed in
E. coli. It shows a high degree of homology to ClpB, a member of the Clp family of proteases and was immunologically reactive with the rabbit hyperimmune serum against
M. bovis BCG. A positive signal was also obtained with sera of patients with tuberculosis. This antigen is a previously unrecognized target of the human immune response to mycobacteria. |
| doi_str_mv | 10.1016/S0962-8479(98)80006-3 |
| format | article |
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M. bovis BCG cosmid library in
M. smegmatis with a hyperimmune rabbit anti-BCG serum.
Cross-reactive antibodies interfere with the detection of BCG specific antigens in
M. smegmatis culture filtrates. We, therefore, screened parallel western blots with serum adsorbed with a
M. smegmatis cell lysate and unadsorbed serum. Comparison of the western blots allowed distinction between BCG specific and cross-reactive
M. smegmatis antigens. Thirty-one cosmids expressed BCG specific antigens. One of them, a hitherto undescribed 100 kDa antigen was subcloned, sequenced and expressed in
E. coli. It shows a high degree of homology to ClpB, a member of the Clp family of proteases and was immunologically reactive with the rabbit hyperimmune serum against
M. bovis BCG. A positive signal was also obtained with sera of patients with tuberculosis. This antigen is a previously unrecognized target of the human immune response to mycobacteria.</description><identifier>ISSN: 0962-8479</identifier><identifier>EISSN: 1532-219X</identifier><identifier>DOI: 10.1016/S0962-8479(98)80006-3</identifier><identifier>PMID: 9692182</identifier><language>eng</language><publisher>Scotland: Elsevier Ltd</publisher><subject>Animals ; Antibody Formation ; Antigens, Bacterial - genetics ; Antigens, Bacterial - isolation & purification ; Base Sequence ; Blotting, Western ; Endopeptidase Clp ; Escherichia coli - immunology ; Escherichia coli Proteins ; Gene Library ; Heat-Shock Proteins - genetics ; Humans ; Molecular Sequence Data ; Mycobacterium - genetics ; Mycobacterium - immunology ; Mycobacterium bovis - genetics ; Mycobacterium bovis - immunology ; Rabbits ; Tuberculosis, Pulmonary - immunology</subject><ispartof>Tubercle and lung disease, 1998-01, Vol.78 (2), p.133-141</ispartof><rights>1998 Harcourt Brace & Co. Ltd All rights reserved</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c339t-5496bf5ce07acf477d95f4398c26bfbad8428ca4cae4f21d899fa899b9f64f73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9692182$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bona, M.</creatorcontrib><creatorcontrib>Nayak, R.</creatorcontrib><creatorcontrib>Wu, M.</creatorcontrib><creatorcontrib>Mincek, M.</creatorcontrib><creatorcontrib>Ellner, J.J.</creatorcontrib><title>Immunological screening of a genomic M. bovis BCG library expressed in M. smegmatis and identification of the M. bovis BCG analog of ClpB</title><title>Tubercle and lung disease</title><addtitle>Tuber Lung Dis</addtitle><description>This work describes the screening of a
M. bovis BCG cosmid library in
M. smegmatis with a hyperimmune rabbit anti-BCG serum.
Cross-reactive antibodies interfere with the detection of BCG specific antigens in
M. smegmatis culture filtrates. We, therefore, screened parallel western blots with serum adsorbed with a
M. smegmatis cell lysate and unadsorbed serum. Comparison of the western blots allowed distinction between BCG specific and cross-reactive
M. smegmatis antigens. Thirty-one cosmids expressed BCG specific antigens. One of them, a hitherto undescribed 100 kDa antigen was subcloned, sequenced and expressed in
E. coli. It shows a high degree of homology to ClpB, a member of the Clp family of proteases and was immunologically reactive with the rabbit hyperimmune serum against
M. bovis BCG. A positive signal was also obtained with sera of patients with tuberculosis. This antigen is a previously unrecognized target of the human immune response to mycobacteria.</description><subject>Animals</subject><subject>Antibody Formation</subject><subject>Antigens, Bacterial - genetics</subject><subject>Antigens, Bacterial - isolation & purification</subject><subject>Base Sequence</subject><subject>Blotting, Western</subject><subject>Endopeptidase Clp</subject><subject>Escherichia coli - immunology</subject><subject>Escherichia coli Proteins</subject><subject>Gene Library</subject><subject>Heat-Shock Proteins - genetics</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><subject>Mycobacterium - genetics</subject><subject>Mycobacterium - immunology</subject><subject>Mycobacterium bovis - genetics</subject><subject>Mycobacterium bovis - immunology</subject><subject>Rabbits</subject><subject>Tuberculosis, Pulmonary - immunology</subject><issn>0962-8479</issn><issn>1532-219X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><recordid>eNqFkc1O3DAUha2qiA60j4DkVVUWAdtxHHtVdUblRwKxKIvuLMe5nrpK7KmdQfQReGscZoTEio0t3fPdc6VzEDqh5IwSKs5_ESVYJXmrvil5Kgkhoqo_oAVtalYxqn5_RItX5BM6yvlvYRgX_BAdKqEYlWyBnq7HcRviENfemgFnmwCCD2scHTZ4DSGO3uLbM9zFB5_xcnWJB98lk_5jeNwkyBl67MNM5BHWo5kKZUKZ9RAm74rr5GOY7aY_8NbIBFPuztJq2Cw_owNnhgxf9v8xur_4eb-6qm7uLq9XP24qW9dqqhquROcaC6Q11vG27VXjeK2kZWXemV5yJq3h1gB3jPZSKWfK0yknuGvrY_R1Z7tJ8d8W8qRHny0MgwkQt1m3StASDX8XpIKzmlFSwGYH2hRzTuD0JvmxJKQp0XNV-qUqPfegldQvVem67J3sD2y7EfrXrX03Rf--06Gk8eAh6Ww9BAu9T2An3Uf_zoVn25Oj3g</recordid><startdate>19980101</startdate><enddate>19980101</enddate><creator>Bona, M.</creator><creator>Nayak, R.</creator><creator>Wu, M.</creator><creator>Mincek, M.</creator><creator>Ellner, J.J.</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>19980101</creationdate><title>Immunological screening of a genomic M. bovis BCG library expressed in M. smegmatis and identification of the M. bovis BCG analog of ClpB</title><author>Bona, M. ; Nayak, R. ; Wu, M. ; Mincek, M. ; Ellner, J.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-5496bf5ce07acf477d95f4398c26bfbad8428ca4cae4f21d899fa899b9f64f73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Animals</topic><topic>Antibody Formation</topic><topic>Antigens, Bacterial - genetics</topic><topic>Antigens, Bacterial - isolation & purification</topic><topic>Base Sequence</topic><topic>Blotting, Western</topic><topic>Endopeptidase Clp</topic><topic>Escherichia coli - immunology</topic><topic>Escherichia coli Proteins</topic><topic>Gene Library</topic><topic>Heat-Shock Proteins - genetics</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><topic>Mycobacterium - genetics</topic><topic>Mycobacterium - immunology</topic><topic>Mycobacterium bovis - genetics</topic><topic>Mycobacterium bovis - immunology</topic><topic>Rabbits</topic><topic>Tuberculosis, Pulmonary - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bona, M.</creatorcontrib><creatorcontrib>Nayak, R.</creatorcontrib><creatorcontrib>Wu, M.</creatorcontrib><creatorcontrib>Mincek, M.</creatorcontrib><creatorcontrib>Ellner, J.J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Tubercle and lung disease</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bona, M.</au><au>Nayak, R.</au><au>Wu, M.</au><au>Mincek, M.</au><au>Ellner, J.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immunological screening of a genomic M. bovis BCG library expressed in M. smegmatis and identification of the M. bovis BCG analog of ClpB</atitle><jtitle>Tubercle and lung disease</jtitle><addtitle>Tuber Lung Dis</addtitle><date>1998-01-01</date><risdate>1998</risdate><volume>78</volume><issue>2</issue><spage>133</spage><epage>141</epage><pages>133-141</pages><issn>0962-8479</issn><eissn>1532-219X</eissn><abstract>This work describes the screening of a
M. bovis BCG cosmid library in
M. smegmatis with a hyperimmune rabbit anti-BCG serum.
Cross-reactive antibodies interfere with the detection of BCG specific antigens in
M. smegmatis culture filtrates. We, therefore, screened parallel western blots with serum adsorbed with a
M. smegmatis cell lysate and unadsorbed serum. Comparison of the western blots allowed distinction between BCG specific and cross-reactive
M. smegmatis antigens. Thirty-one cosmids expressed BCG specific antigens. One of them, a hitherto undescribed 100 kDa antigen was subcloned, sequenced and expressed in
E. coli. It shows a high degree of homology to ClpB, a member of the Clp family of proteases and was immunologically reactive with the rabbit hyperimmune serum against
M. bovis BCG. A positive signal was also obtained with sera of patients with tuberculosis. This antigen is a previously unrecognized target of the human immune response to mycobacteria.</abstract><cop>Scotland</cop><pub>Elsevier Ltd</pub><pmid>9692182</pmid><doi>10.1016/S0962-8479(98)80006-3</doi><tpages>9</tpages></addata></record> |
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| ispartof | Tubercle and lung disease, 1998-01, Vol.78 (2), p.133-141 |
| issn | 0962-8479 1532-219X |
| language | eng |
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| source | ScienceDirect Freedom Collection |
| subjects | Animals Antibody Formation Antigens, Bacterial - genetics Antigens, Bacterial - isolation & purification Base Sequence Blotting, Western Endopeptidase Clp Escherichia coli - immunology Escherichia coli Proteins Gene Library Heat-Shock Proteins - genetics Humans Molecular Sequence Data Mycobacterium - genetics Mycobacterium - immunology Mycobacterium bovis - genetics Mycobacterium bovis - immunology Rabbits Tuberculosis, Pulmonary - immunology |
| title | Immunological screening of a genomic M. bovis BCG library expressed in M. smegmatis and identification of the M. bovis BCG analog of ClpB |
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