Cloning of the replication and incompatibility regions of a plasmid derived from Rts1 |
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Authors: | Yoshifumi Itoh Yoshiyuki Kamio Yuko Furuta Yoshiro Terawaki |
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Affiliation: | Department of Bacteriology, Shinshu University School of Medicine, Asahi, 3-1-1, Matsumoto 390, Japan |
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Abstract: | A replication region, consisting of a 1.1-megadalton (Md) EcoRI/HindIII fragment, was isolated from an Rts1 derivative plasmid. This 1.1-Md fragment, designated as mini-Rts1, was ligated to either pBR322 or a nonreplicating DNA fragment specifying a drug resistance, and its replication properties were investigated. The mini-Rts1 plasmid was cured at a high frequency at 42 °C, while it was maintained stably at 37 °C despite it existed in low copy number. These behaviors are quite similar to those of Rts1. By dissecting the pBR322:mini-Rts1 chimeric plasmid with AccI endonuclease, an inc region of 0.34 Md in size was cloned, which expressed incompatibility toward Rts1. Proteins encoded on the mini-Rts1 genome were examined in the minicell system, and one specific product of 35,000 daltons in molecular weight was identified. Any polypeptides specific for the 0.34-Md inc+ region within mini-Rts1 were not detected. |
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Keywords: | To whom correspondence should be addressed. |
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