SUMMARY: A sample of colonies from the Clarke-Carbon Co1E1- DNA plasmid gene bank was screened by conjugation for complementation of the lipoamide dehydrogenase lesion of a deletion strain lacking all components of the pyruvate dehydrogenase complex, Δ(). Two Co1E1- hybrid plasmids were identified: pGS2 (Co1E1- ; 24 kb) and pGS5 (Co1E1- ; 14 kb). Enzymological studies confirmed that pGS2 expressed all the activities of the pyruvate dehydrogenase complex, whereas pGS5 expressed the lipoamide dehydrogenase and acetyltransferase activities (the latter from a ColE1 promoter). These and other plasmids were used to construct a 47-site (15 enzymes) restriction map for a 24.2 kb segment of bacterial DNA in the region. A futher 13 sites (six enzymes) were defined in a 5.4 kb sub-segment containing the gene. λ phage derivatives containing specific fragments were constructed and used in transduction studies which located the and genes in a 7.78 kb sub-segment flanked by and sites.


Article metrics loading...

Loading full text...

Full text loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error