New shuttle vectors for Escherichia coli and Bacillus subtilis. IV The nucleotide sequence of pHY300PLK and some properties in relation to transformation.

Abstract

The whole nucleotide sequence of pHY300PK (Ishiwa and Shibahara 1985a) has been determined. pHY300PLK consists of 4870 bp and contains an RNA primer for ori-177 and three open reading frames corresponding to TcR, ApR and Rep-.alpha.1. Twenty-three unique restriction endonuclease recognition sites were found. We have confirmed that ORF .alpha.1 of pHY300PLK is the plasmid replication gene. A rescue phenomenon has been found with respect to multiple infection by pHY300.2PLK (rep-) and pUB110. A series of experiments indicated that the oligomer form, pHY300.2PLK, was rescued by the monomeric form, pUB110. Additionally, some useful information for using pHY300PLK as a shuttle vector in molecular cloning studies is presented.