ISSN:
1432-0991
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Medicine
Notes:
Abstract A 9.5-kb shuttle vector capable of replication and selection in both Escherichia coli and Butyrivibrio fibrisolvens was constructed. Plasmid pUC118 provided replication functions and ampicillin resistance selection in E. coli. In B. fibrisolvens, replication was controlled by the native plasmid pRJF1 from strain OB156, and selectability was provided by a 3.5-kb fragment of plasmid pAMβ1 containing the erythromycin resistance gene. Optimum conditions for transformation were 15 kV/cm, 2 h recovery, and plating in an agar overlay on medium containing 10 μg erythromycin/ml. Maximum efficiency was 1.1×105 transformants per μg plasmid DNA (average 3×104), and restriction mechanisms reduced efficiency by a factor of 2×102. Nonselective growth for 200 generations gave no measurable loss of plasmid.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00294191
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