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The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing.
Gene 1988; 68(1):139-49GENE

Abstract

A series of nic- cloning vectors have been constructed analogous to the pUC plasmids but which are smaller in size and carry more extensive polylinker regions within the lacZ' gene. The vectors pMTL20 and pMTL21 carry six additional sites (AatII, MluI, NcoI, BglII, XhoI and StuI) to those present in pUC18 and pUC19, while pMTL22 and -23 possess eleven new cloning sites (ActII, MluI, NcoI, BglII, XhoI, StuI, NaeI, EcoRV, ClaI, NdeI and NruI). More importantly, the relative order of the restriction sites within the polylinker of these latter vectors has been totally rearranged, relative to pUC18 and pUC19, to facilitate the conversion of DNA fragments with incompatible ends to fragments with compatible termini. The availability of such DNA fragments is a crucial requirement when M13 templates are generated for dideoxy sequencing by the sonication procedure. Derivatives of these vectors have also been constructed which demonstrate improved segregational stability by incorporation of the pSC101 par locus. During the construction of these new vectors data were obtained which demonstrated that the pUC and pMTL plasmids contain a previously unreported single base pair difference within the RNA I/RNA II region (compared to pBR322) responsible for a three-fold increase in plasmid copy number. The pUC and pMTL plasmids were also shown to be functionally nic-, thus affording the lowest categorisation in genetic manipulation experiments.

Authors+Show Affiliations

Microbial Technology Laboratory, PHLS Centre for Applied Microbiology and Research, Salisbury, Wiltshire, U.K.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

2851488

Citation

Chambers, S P., et al. "The pMTL Nic- Cloning Vectors. I. Improved pUC Polylinker Regions to Facilitate the Use of Sonicated DNA for Nucleotide Sequencing." Gene, vol. 68, no. 1, 1988, pp. 139-49.
Chambers SP, Prior SE, Barstow DA, et al. The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene. 1988;68(1):139-49.
Chambers, S. P., Prior, S. E., Barstow, D. A., & Minton, N. P. (1988). The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene, 68(1), pp. 139-49.
Chambers SP, et al. The pMTL Nic- Cloning Vectors. I. Improved pUC Polylinker Regions to Facilitate the Use of Sonicated DNA for Nucleotide Sequencing. Gene. 1988 Aug 15;68(1):139-49. PubMed PMID: 2851488.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. AU - Chambers,S P, AU - Prior,S E, AU - Barstow,D A, AU - Minton,N P, PY - 1988/8/15/pubmed PY - 1988/8/15/medline PY - 1988/8/15/entrez SP - 139 EP - 49 JF - Gene JO - Gene VL - 68 IS - 1 N2 - A series of nic- cloning vectors have been constructed analogous to the pUC plasmids but which are smaller in size and carry more extensive polylinker regions within the lacZ' gene. The vectors pMTL20 and pMTL21 carry six additional sites (AatII, MluI, NcoI, BglII, XhoI and StuI) to those present in pUC18 and pUC19, while pMTL22 and -23 possess eleven new cloning sites (ActII, MluI, NcoI, BglII, XhoI, StuI, NaeI, EcoRV, ClaI, NdeI and NruI). More importantly, the relative order of the restriction sites within the polylinker of these latter vectors has been totally rearranged, relative to pUC18 and pUC19, to facilitate the conversion of DNA fragments with incompatible ends to fragments with compatible termini. The availability of such DNA fragments is a crucial requirement when M13 templates are generated for dideoxy sequencing by the sonication procedure. Derivatives of these vectors have also been constructed which demonstrate improved segregational stability by incorporation of the pSC101 par locus. During the construction of these new vectors data were obtained which demonstrated that the pUC and pMTL plasmids contain a previously unreported single base pair difference within the RNA I/RNA II region (compared to pBR322) responsible for a three-fold increase in plasmid copy number. The pUC and pMTL plasmids were also shown to be functionally nic-, thus affording the lowest categorisation in genetic manipulation experiments. SN - 0378-1119 UR - https://www.unboundmedicine.com/medline/citation/2851488/The_pMTL_nic__cloning_vectors__I__Improved_pUC_polylinker_regions_to_facilitate_the_use_of_sonicated_DNA_for_nucleotide_sequencing_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0378-1119(88)90606-3 DB - PRIME DP - Unbound Medicine ER -