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Characterization of DNA synthesis and chloroplast DNA replication initiation in a Petunia hybrida chloroplast lysate system |
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| Authors: | Jan M. de Haas Ad J. Kool Nico Overbeeke Wieger van Brug H. John J. Nijkamp |
| Affiliation: | (1) Department of Genetics, Vrije Universiteit, De Boelelaan 1087, NL-1081HV Amsterdam, The Netherlands;(2) Present address: Zaadunie B.V., Plant Biotechnology Division, P.O. Box 26, NL-1600 AA Enkhuizen, The Netherlands;(3) Present address: Unilever Research Laboratory, Olivier van Noortlaan 120, NL-3133 AT Vlaardingen, The Netherlands |
| Abstract: | Summary An in vitro chloroplast DNA synthesizing lysate system prepared from purified chloroplasts of Petunia hybrida leaves has been developed. Both co-isolated endogeneous chloroplast (cp)DNA and externally added DNA can be used as DNA templates in the system. The system contains a  -like DNA polymerase as determined by using DNA polymerase-specific inhibitors and synthetic templates. The molecular weight of this enzyme is about 85 kd. Part of the DNA synthesizing activity is repair synthesis. When a chimaeric plasmid containing a fragment with a potential cpDNA replication origin is used as a template (pPCY62), specific initiation of DNA synthesis is observed on this fragment which strongly suggests that the in vitro chloroplast lysate system is also capable of replication initiation. |
| Keywords: | ARS Chloroplast Petunia hybrida Replication |
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