PSET152 - Revision history

Matt Hutchings: /* References */

2026-10-10T09:28:37Z

References

← Older revision		Revision as of 09:28, 10 October 2026
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	==References==		==References==

			M. Bierman, R. Logan, K. O'Brien, E.T. Seno, R. Nagaraja Rao, B.E. Schoner (1992). Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43-49.

			Abstract
			We have constructed cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. All vectors contain the 760-bp oriT fragment from the IncP plasmid, RK2. Transfer functions need to be supplied in trans by the E. coli donor strain. We have incorporated into these vectors selectable antibiotic-resistance markers (AmR, ThR, SpR) that function in Streptomyces spp. and other features that should allow for: (i) integration via homologous recombination between cloned DNA and the Streptomyces spp. chromosome, (ii) autonomous replication, or (iii) site-specific integration at the bacteriophage φC31 attachment site. Shuttle cosmids for constructing genomic libraries and bacteriophage P1 cloning vector capable of accepting approx. 100-kb fragments are also described. A simple mating procedure has been developed for the conjugal transfer of these vectors from E. coli to Streptomyces spp. that involves plating of the donor strain and either germinated spores or mycelial fragments of the recipient strain. We have shown that several of these vectors can be introduced into Streptomyces fradiae, a strain that is notoriously difficult to transform by PEG-mediated protoplast transformation.


			https://www.sciencedirect.com/science/article/abs/pii/0378111992906272

Matt Hutchings: /* Features */

2026-10-10T09:26:06Z

Features

← Older revision		Revision as of 09:26, 10 October 2026
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	==Features==		==Features==

	• Backbone: Derived from a standard E. coli plasmid vector (~~such as~~ pBR322/pUC lineage) containing an E. coli replication origin (ori) and an origin of transfer (oriT) derived from a broad-host-range plasmid ~~like~~ RK2 to allow conjugal transfer.		• Backbone: Derived from a standard E. coli plasmid vector (pBR322/pUC lineage) containing an E. coli replication origin (ori) and an origin of transfer (oriT) derived from broad-host-range plasmid RK2 to allow conjugal transfer.

	• Integrative Machinery: Incorporates the attachment site (attP) and the integrase gene (int) from the temperate actinophage φC31.		• Integrative Machinery: Incorporates the attachment site (attP) and the integrase gene (int) from the temperate actinophage φC31. Integrates into the φC31 att site on the Streptomyces genome.

	• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.		• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.

Matt Hutchings at 09:24, 10 October 2026

2026-10-10T09:24:57Z

← Older revision		Revision as of 09:24, 10 October 2026
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	{{DISPLAYTITLE: pSET152}}		{{DISPLAYTITLE: pSET152}}



	==Features==		==Features==
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	• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.		• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.


	==History==		==History==

Matt Hutchings at 09:24, 10 October 2026

2026-10-10T09:24:33Z

← Older revision		Revision as of 09:24, 10 October 2026
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	~~Key Components & Origin~~
			==Features==

	• Backbone: Derived from a standard E. coli plasmid vector (such as pBR322/pUC lineage) containing an E. coli replication origin (ori) and an origin of transfer (oriT) derived from a broad-host-range plasmid like RK2 to allow conjugal transfer.		• Backbone: Derived from a standard E. coli plasmid vector (such as pBR322/pUC lineage) containing an E. coli replication origin (ori) and an origin of transfer (oriT) derived from a broad-host-range plasmid like RK2 to allow conjugal transfer.
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	• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.		• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.

	~~==Features==~~

	~~Apramycin resistance marker allows for selection of the plasmid after conjkugation from E. coli~~		==History==

	~~==History==~~		The pSET152 plasmid was constructed and published by Bierman et al. in 1992 as an E. coli–Streptomyces shuttle and integrative vector

	==Map==		==Map==

Matt Hutchings: /* Use */

2026-10-10T09:23:44Z

Use

← Older revision		Revision as of 09:23, 10 October 2026
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	~~==Use==~~		Key Components & Origin

	~~Integrating genes in trans into~~ the ~~phi C31 phage~~ attachment site. ~~This can be used for complementation of mutants with wild-type alleles under native~~, ~~constitutive or inducible promoters or alleles encoding Flag-tagged proteins for ChIP-seq~~ and ~~co-IP~~.		• Backbone: Derived from a standard E. coli plasmid vector (such as pBR322/pUC lineage) containing an E. coli replication origin (ori) and an origin of transfer (oriT) derived from a broad-host-range plasmid like RK2 to allow conjugal transfer.

			• Integrative Machinery: Incorporates the attachment site (attP) and the integrase gene (int) from the temperate actinophage φC31.

			• Selection Marker: Carries the aac(3)IV gene, which confers resistance to apramycin in both E. coli and Streptomyces.

	==Features==		==Features==

Matt Hutchings: /* Features */

2026-10-10T09:22:37Z

Features

← Older revision		Revision as of 09:22, 10 October 2026
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	==Features==		==Features==

			Apramycin resistance marker allows for selection of the plasmid after conjkugation from E. coli

	==History==		==History==

Matt Hutchings: /* Use */

2026-10-10T09:22:03Z

Use

← Older revision		Revision as of 09:22, 10 October 2026
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	==Use==		==Use==

	Integrating genes in trans into the phi C31 ~~ohage~~ attachment site		Integrating genes in trans into the phi C31 phage attachment site. This can be used for complementation of mutants with wild-type alleles under native, constitutive or inducible promoters or alleles encoding Flag-tagged proteins for ChIP-seq and co-IP.

	==Features==		==Features==

Matt Hutchings: /* Use */

2026-10-10T09:21:03Z

Use

← Older revision		Revision as of 09:21, 10 October 2026
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	==Use==		==Use==

			Integrating genes in trans into the phi C31 ohage attachment site

	==Features==		==Features==

Sam Prudence at 16:39, 8 August 2019

2019-08-08T16:39:18Z

← Older revision		Revision as of 16:39, 8 August 2019
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			{{DISPLAYTITLE: pSET152}}


	==Use==		==Use==

Sam Prudence at 16:22, 8 August 2019

2019-08-08T16:22:33Z

← Older revision		Revision as of 16:22, 8 August 2019
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	~~{{DISPLAYTITLE: pSET152}}~~		==Use==

	=~~pSET152~~=		==Features==


	==~~Background~~==		==History==

	==~~Construction~~==		==Map==
			[[File:pSET152.png]]
	~~==Uses==~~

	~~==Characteristics==~~

	~~==Map and Sequence==~~

	~~[[File:pSET152.png]]~~		==Sequence links==

	==References==		==References==