Cloning, sequencing and identification of replication origin of Rhodococcus linear plasmid pNSL

Abstract:

Abstract: 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 4.1040. [Conclusion] Cloning and sequencing of Rhodococcus linear plasmid pNSL1, and identification of its replication origin.