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[Cloning, sequencing and identification of replication origin of Rhodococcus linear plasmid pNSL1]
Yingmin Zhu1, Mingxuan Xu, Meijuan Shen
1Key Laboratory of Synthetic Biology, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China. zoymin@gmail.com
Unlabelled:
Two linear plasmids, pNSL1 and pNSL1, were detected from Rhodocuccus sp. NS1.
Objective:
Cloning, sequencing and identification of replication origin of the Rhodococcus linear plasmid pNSL1.
Methods:
Large amount of linear plasmid DNA was recovered from pulsed-field gels for shotgun-cloning and sequencing, and identification of its replication locus.
Results:
The complete nucleotide sequence of pNSL1 consisted of 117252 bp, including the conserved 1282-bp telomere sequences among Rhodococcus linear plasmids. pNSL1 encoded 103 open reading frames, including functions of replication, maintenance and transfer etc. A locus, pNSL1. 038 and upstream 767-bp non-coding sequence, was identified for autonomous replication by cloning in an E. coli vector and introduced by electroporation into Nocardia coralline 4. 1037.
Conclusion:
Cloning and sequencing of Rhodococcus linear plasmid pNSL1, and identification of its replication origin.
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