Development of a reporter system for the study of gene expression for solvent production in Clostridium beijerinckii NRRL B592 and Clostridium acetobutylicum ATCC 824

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dc.contributor.authorLi, Guang-Shanen
dc.contributor.committeechairChen, Jiann-Shinen
dc.contributor.committeememberKrieg, Noel R.en
dc.contributor.committeememberNewton, William E.en
dc.contributor.committeememberDean, Dennis R.en
dc.contributor.committeememberLarson, Timothy J.en
dc.contributor.departmentBiochemistry and Anaerobic Microbiologyen
dc.date.accessioned2014-03-14T20:17:53Zen
dc.date.adate1998-12-11en
dc.date.available2014-03-14T20:17:53Zen
dc.date.issued1998-09-23en
dc.date.rdate1999-12-11en
dc.date.sdate1998-11-03en
dc.description.abstractTo study the regulation of gene expression, a good reporter system is very useful. The lack of a good reporter system for the solvent-producing clostridia hindered the progress of research in this area. The objective of this study was to develop a reporter system to facilitate the elucidation of the control mechanism for the expression of solvent-producing genes. A potential reporter gene was found in Clostridium beijerinckii NRRL B593, which contains an adh gene encoding a primary-secondary alcohol dehydrogenase and this adh gene is not present in Clostridium acetobutylicum ATCC 824 and Clostridium beijerinckii NRRL B592. The adh gene was cloned into the E. coli -Clostridium shuttle vectors to generate plasmids. An electro-transformation procedure was developed for C. beijerinckii NRRL B592. Shuttle plasmids were transformed into C. beijerinckii NRRL B592 or C. acetobutylicum ATCC 824. The copy number of the plasmids in C. beijerinckii was 4. Isopropanol production suggested that the adh gene was expressed in transformants of C. acetobutylicum ATCC 824 and C. beijerinckii NRRL B592. Northern analysis indicated that the expression of the adh gene was regulated at the transcriptional level in the transformants of C. beijerinckii. The transcriptional start site for the adh gene was identified by the primer extension method. A promoter-probing vector was constructed and tested with the promoter from the ferredoxin(fer) gene. The expression of the adh gene under the control of the fer promoter was at a low and similar level during acidogenesis and solventogenesis. The expression pattern of the adh gene under the control of the promoter of the adh gene differed from that under the control of the promoter of the fer gene.en
dc.description.degreePh. D.en
dc.identifier.otheretd-110298-180740en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-110298-180740/en
dc.identifier.urihttp://hdl.handle.net/10919/29439en
dc.publisherVirginia Techen
dc.relation.haspartETDall.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectadhen
dc.subjectreporter systemen
dc.subjectanaerobeen
dc.subjectsolventogenesisen
dc.subjectClostridiumen
dc.titleDevelopment of a reporter system for the study of gene expression for solvent production in Clostridium beijerinckii NRRL B592 and Clostridium acetobutylicum ATCC 824en
dc.typeDissertationen
thesis.degree.disciplineBiochemistry and Anaerobic Microbiologyen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.leveldoctoralen
thesis.degree.namePh. D.en

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