The origin of replication of plasmid p15A and comparative studies on the nucleotide sequences around the origin of related plasmids

Shibahara

1985

A small plasmid pHY163PLK (2519 bp) was constructed from the shuttle vector pHY300PLK. pHY163PLK contains the replication origin of pACYC-177, the tetracycline resistance gene (tetal) of pAMal derived from Streptococcus faecalis and eight continuous unique cloning sites of a polylinker region (EcoRI, SmaI, BamHI, SalI, PstI, BglII, XbaI and HindIII), pHY163PLK, a copy-number mutant, has one base substitution in RNA I region compared with that of p15A which is the ancestor of ori-pHY163PLK.The tetal, which codes 458 amino acids (1374 bp), is highly homologous to the TcR genes of other Gram-positive bacteria but completely different from the TcR gene encoded by pBR322 in Gram-negative bacteria. The regulatory region of tetal (constitutive type) has an insertion of seven base pairs in the leader peptide region of a related inducible TcR

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

New shuttle vectors for Escherichia coli and Bacillus subtilis. III. Nucleotide sequence analysis of tetracycline resistance gene of pAM.ALPHA.1 and ori-177.

Shibahara

1985

“…Actually, the nucleotide sequence analysis of pHY300PLK (H. I., in preparation) has revealed that there is a single base substitution in the regulatory RNA I when compared with the on-p15A (Selzer et al 1983). Note that pACYC177 was derived from the pl5A replicon (Chang and Cohen 1978).…”

Section: Resultsmentioning

confidence: 99%

New shuttle vectors for Escherichia coli and Bacillus subtilis. II. Plasmid pHY300PLK, a multipurpose cloning vector with a polylinker, derived from pHY460.

Shibahara

1985

In vivo and in vitro recombination techniques were used to construct a new cloning vector, pHY300PLK (4.7 kb) from the shuttle vector pHY460 (7 kb). The newly derived shuttle vector can replicate and express the tetracycline resistance gene (TcR) in both Escherichia coli and Bacillus subtilis. pHY300PLK contains the TcR gene, the ampicillin resistance gene (ApR), two replication origins for EK coli and B. subtilis and a polylinker derived from ~AN7. The unique cloning sites are BalI,

“…Therefore, complementation did not occur between the two replicons, nor was a cointegrate The RNA primer for on-177 in pHY300,PLK corresponds to p15A. Each of the replication origins, on-177 derived from pACYC177 (Selzer et al 1983), and on-al derived from pAMal (Scheer-Abramowitz et al 1981) is also indicated with open arrows, respectively. Fig.…”

Section: Methodsmentioning

confidence: 99%

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

Shibahara-Sone

1986

The whole nucleotide sequence of pHY300PLK (Ishiwa and Shibahara 1985a) has been determined, pHY300PLK consists of 4870 by and contains an RNA primer for on-177 and three open reading frames corresponding to TcR, ApR and Rep-al. Twenty-three unique restriction endonuclease recognition sites were found. We have confirmed that ORF al 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.