Some Aspects of E. coli Promoter Evolution Observed in a Molecular Evolution Experiment |
| |
Authors: | Shumo Liu Albert Libchaber |
| |
Institution: | (1) NEC Laboratories America, 4 Independence Way, Princeton, NJ 08540, USA;(2) Rockefeller University, 1230 York Avenue, New York, NY 10021, USA;(3) Department of Physics, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA |
| |
Abstract: | We devised a molecular evolution procedure to evolve E. coli promoter sequences and applied it to observe an arbitrary, nonfunctional sequence evolving into functional promoters. In
the experiments, DNA sequence variations were generated with error-prone PCR and were inserted in the promoter region of the
cat (chloramphenicol acetyl transferase) gene on a plasmid. Upon transforming the cells, functional promoters on the plasmid
were selected according to the chloramphenicol resistance. Within a few cycles of mutation-selection, promoters emerged, and
the sequences converged into a small number of groups. In the process, the extended minus 10 type of promoters emerged quickly,
and small deletions were often involved in adjusting the length between the −35 and the −10 elements. Our results also suggest
a possible selection for promoter stability against mutation.
Reviewing Editior: Dr. Laura Landweber] |
| |
Keywords: | Promoter Mutagenesis PCR Antibiotic selection Mutation frequency Extended minus 10 |
本文献已被 PubMed SpringerLink 等数据库收录! |
|