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Mapping of RP4 plasmid using deletion mutants of pAS8 hybrid (RP4--ColE1).
Mol Gen Genet. 1978 Oct 24; 165(3):331-41.MG

Abstract

We used the hybrid plasmid pAS8 in order to conduct the genetic analysis of RP4 plasmid. The presence of two replicons in the hybrid plasmid permitted to expand the spectrum of deletion mutants of RP4 isolated, which are capable to autonomous replication. The shortening of the hybrid plasmid was achieved by P22 transduction, by induction of deletion mutants using mitomycin C, as well as by seletion of Tra- mutants on the basis of resistance of cells to P-specific phages. These techniques have lead to isolation of clones possessing different combinations of plasmid resistance determinants. Comparison of phenotypic characteristics of deletion plasmids pAS9, pAS10, pAS11, pAS12 and pAS10-2 permitted to propose the map for pAS8 plasmid with the following sequence of markers: tra--kam--ColE1--amp--tet... Heteroduplex analysis of deletion mutants of pAS8 permitted to construct a physical map and to elaborate in greater detail the functional map of RP4 plasmid. The correlation between the ability of mutants to replicate in polA(TS) strain at nonpermissive conditions and the length of the deleted segment permitted to map rep genes of RP4 on a region with coordinates 9.8--17.3 kb. A relationship between the manifestation of incompatibility of mutants with Inc P-1 plasmids and the length of deletions points out that inc genes are located on DNA region with coordinates 2.1--9.8 kb. The analysis of replication of deletion mutants and the manifestation of incompatibility just as of the data about the size of appropriate deletions permitted to make the conclusion about the functional and genetic independence of the replication control and incompatibility control in RP4 plasmid.

Authors

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Pub Type(s)

Journal Article

Language

eng

PubMed ID

368565

Citation

Sakanyan, V A., et al. "Mapping of RP4 Plasmid Using Deletion Mutants of pAS8 Hybrid (RP4--ColE1)." Molecular & General Genetics : MGG, vol. 165, no. 3, 1978, pp. 331-41.
Sakanyan VA, Yakubov LZ, Alikhanian SI, et al. Mapping of RP4 plasmid using deletion mutants of pAS8 hybrid (RP4--ColE1). Mol Gen Genet. 1978;165(3):331-41.
Sakanyan, V. A., Yakubov, L. Z., Alikhanian, S. I., & Stepanov, A. I. (1978). Mapping of RP4 plasmid using deletion mutants of pAS8 hybrid (RP4--ColE1). Molecular & General Genetics : MGG, 165(3), 331-41.
Sakanyan VA, et al. Mapping of RP4 Plasmid Using Deletion Mutants of pAS8 Hybrid (RP4--ColE1). Mol Gen Genet. 1978 Oct 24;165(3):331-41. PubMed PMID: 368565.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mapping of RP4 plasmid using deletion mutants of pAS8 hybrid (RP4--ColE1). AU - Sakanyan,V A, AU - Yakubov,L Z, AU - Alikhanian,S I, AU - Stepanov,A I, PY - 1978/10/24/pubmed PY - 1978/10/24/medline PY - 1978/10/24/entrez SP - 331 EP - 41 JF - Molecular & general genetics : MGG JO - Mol. Gen. Genet. VL - 165 IS - 3 N2 - We used the hybrid plasmid pAS8 in order to conduct the genetic analysis of RP4 plasmid. The presence of two replicons in the hybrid plasmid permitted to expand the spectrum of deletion mutants of RP4 isolated, which are capable to autonomous replication. The shortening of the hybrid plasmid was achieved by P22 transduction, by induction of deletion mutants using mitomycin C, as well as by seletion of Tra- mutants on the basis of resistance of cells to P-specific phages. These techniques have lead to isolation of clones possessing different combinations of plasmid resistance determinants. Comparison of phenotypic characteristics of deletion plasmids pAS9, pAS10, pAS11, pAS12 and pAS10-2 permitted to propose the map for pAS8 plasmid with the following sequence of markers: tra--kam--ColE1--amp--tet... Heteroduplex analysis of deletion mutants of pAS8 permitted to construct a physical map and to elaborate in greater detail the functional map of RP4 plasmid. The correlation between the ability of mutants to replicate in polA(TS) strain at nonpermissive conditions and the length of the deleted segment permitted to map rep genes of RP4 on a region with coordinates 9.8--17.3 kb. A relationship between the manifestation of incompatibility of mutants with Inc P-1 plasmids and the length of deletions points out that inc genes are located on DNA region with coordinates 2.1--9.8 kb. The analysis of replication of deletion mutants and the manifestation of incompatibility just as of the data about the size of appropriate deletions permitted to make the conclusion about the functional and genetic independence of the replication control and incompatibility control in RP4 plasmid. SN - 0026-8925 UR - https://www.unboundmedicine.com/medline/citation/368565/Mapping_of_RP4_plasmid_using_deletion_mutants_of_pAS8_hybrid__RP4__ColE1__ L2 - https://medlineplus.gov/genesandgenetherapy.html DB - PRIME DP - Unbound Medicine ER -