Article Abstract:
The development of a gene expression reporter system (pHT3) for Clostridium acetobutylicum ATCC 824 using the lacZ gene from Thermoanaerobacterium thermosulfurogenes as the reporter gene is described. Promoters of three key metabolic pathway genes were cloned upstream of the reporter gene in pHT3 to construct vectors for pHT4, pHT5 and pHTA and to test the reporter system. REsults indicate that the beta-galactosidase generated by the lacZ gene from T thermosulfurogenes was stable in the exponential phase of growth.
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Article Abstract:
The heterologous expression of Clostridium acetobutylicum ATCC 824 genes in Escherichia coli for acetone production and the prevention of acetate accumulation is reported. It is the first time that the ability to use clostridial genes in nonclostridial hosts for solvent production is demonstrated. Results also revealed the possibility of acetone-producing E. coli strains of being useful hosts for recombinant protein production so that acetate accumulation can be prevented.
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Article Abstract:
The protection of coresident Escherichia coli-Clostridium acetobutylicum shuttle vectors and their introduction into C. acetobutylicum ATCC 824 by electrotransformation is facilitated by the expression of o3T I methyltransferase gene from a compatible plasmid. The utility of in vivo methylation method for circumventing restriction barriers of mutagens was demmonstrated.
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