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Freeze-dried E. coli containing the plasmid
Comments
Restriction digests of the clone give the following sizes (kb): BamHI/EcoRI--3.2, 3.0, 2.5; EcoRI--8.5; SalI--8.5.
The GenBank accession number for the adh insert is M15394.
Strains containing pLOI297 (ATCC 68239) appear to produce higher levels of ethanol than strains containing pLOI1308-11 (ATCC 68238). ATCC strains 11303 and 15224 transformed with pLOI297 appear to be the best constructs for ethanol production.
The 3.2 kb BamHI/EcoRI fragment is the same as that in pLOI295 (ATCC 68240) and contains the two genes.
Constructed by inserting a 2.6 kb SalI fragment from pCOS2EMBL containing tetR into the SalI site of pLOI295 (ATCC 68240).
The pdc reading frame begins 163 bp downstream of the lac promoter and ends with two stop codons 85 bp upstream of the adhB reading frame. The adhB gene has a single stop codon followed by a palindromic sequence which serves as a terminator.
Expression appears to be regulated by the lac promoter.
References
Ingram LO, et al. Ethanol production by Escherichia coli strains co-expressing Zymomonas pdc and adh genes. US Patent 5,000,000 dated Mar 19 1991
Conway T, et al. Cloning and sequencing of the alcohol dehydrogenase II gene from Zymomonas mobilis. J. Bacteriol. 169: 2591-2597, 1987. PubMed: 3584063
Conway T, et al. Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase. J. Bacteriol. 169: 949-954, 1987. PubMed: 3029037
Disclosure
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC.
