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KMID : 0379119860140040000
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Korean Journal of Mycology
1986 Volume.14 No. 4 p.0 ~ p.0
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2. Construction of a New Vector for the Expression of Foreign Genes in Zymomonas mobilis
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Abstract
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Broad host range plasmids have previously been shown to be suitable as vectors to introduce antibiotic resistance genes into Z. mobilis. However, attempts to use these vectors to carry other genes with enteric promoters and controlling elements have resulted in limited success due to poor expression. Thus we have constructed a promoter cloning vector in a modified pBR327 and used this vector to isolate 12 promoters from Z. mobilis which express various levels of beta-galactosidase in E. coli. Four of these were then subcloned into a modified pRK290 for introduction into Z. mobilis. All expressed beta-galactosidase in Z. mobilis with activities of 100 to 1800 miller units. One of these retained a Bam H1 site into which new genes can be readily inserted immediately downsteam from the Z. mobilis promoter. Genetic traits carried by the modified pRK290 were initially unstable but spontaneous variants were produced during subculture in which the plasmid was resistant to curing at elevated temperature. One of these variants was examined in some detail. The increased stability of this variant appears to result from an alteration in the plasmid rather than a chromosomal mutation or from chromosomal integration.
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