Broad host rangeli plasmid vector pghost5 complete. We have constructed a series of broadhostrange plasmids which use visual screens. Host range host range may be wide or narrow broad host range. The content of the open access version may differ from that of the licensed version. A plasmid is a small, extrachromosomal dna molecule within a cell that is physically separated. The rk2 plasmid is a broad host range plasmid belonging to the incp incompatibility group it is notable for its ability to replicate in a wide variety of singlecelled organisms, which makes it suitable as a genetic engineering tool. Broadhostrange bacterial cloning vector with the puc1819 mcs and a kanamycin resistance gene. Cloning vectors cloning vectors are dna molecules that are used to transport cloned sequences between biological hosts and the test tube. Using broad host range replicons as the basis of cloning, vector development has the advantage that the cloning may be performed by standard techniques in e. It is capable of transfer, replication, and maintenance in most genera of gramnegative bacteria. Pmc free article olsen rh, debusscher g, mccombie wr. Broadhostrange prouser vectors enable fast characterization. It is often used to transfer dna constructs designed in escherichia coli to recipient bacteria, yeast, plants and mammalian cells.
Broad host rangeli plasmid vector pmmb67eh complete. Broad host range plasmids fems microbiology letters. Bacterial conjugation is a process that is mediated either by a direct celltocell junction or by formation of a bridge between the cells. They comprise restrictionnegative host strains of pseudomonas aeruginosa and p. These plasmids are small in size host range cloning vector pbbr1mcs, carrying different antibioticresistance cassettes.
Plasmids bearing the rk2rp4 origin of transfer orit are mostly mobilized using the e. In this study, a small broad host range vector named pbbr1mcs 2, which does not belong to the incq, incw, or incp groups, was studied for the feasibility of its use in conjugative gene transfer. Convenient broadhostrange unstable vectors for studying. Broad host range plasmids summary broad host range plasmids are model systems for exploringreplicationmechanismsindiversebacterial species and can extend our. Get a printable copy pdf file of the complete article 626k, or click on a. A vector is used to amplify a single molecule of dna into many copes. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the.
The supporting information is available free of charge on the acs publications website at doi. We have succeeded in preparing and cloning high quality, high molecular weight metagenomic dna of an average insert size of 110 kb from soil using broad host range bacterial artificial chromosome bac shuttle vectors for expression in grampositive and gramnegative hosts. Download fulltext pdf a transposon vector for broad host range cloning article pdf available in nucleic acids research 1819. Host vector systems have been developed for gene cloning in the metabolically versatile bacterial genus pseudomonas.
Kovach me, elzer ph, hill ds, robertson gt, farris ma, roop rm 2nd, peterson km. In this study, a small broad host range vector named pbbr1mcs2, which does not belong to the incq, incw, or incp groups, was studied for the feasibility of its use in conjugative gene transfer. Pseudomonas putida u ppu, rfr tipo, cect 4848 was from our collection 7. They belong to a family of broad host range cloning and expression vectors pbbr1mcs2 kmr. Here, we introduce a series of new rk2 based broad host range expression plasmids for inducible production of affinity and fluorescence tagged proteins in the cytoplasm and periplasm of a wide range of gram negative hosts which are designed to match the recently suggested modular standard european vector architecture and database. Four new derivatives of the broadhostrange cloning. The sequence was not resequenced but deduced from the pbr322 sequence. Get a printable copy pdf file of the complete article 1. Construction of broadhostrange plasmid vectors for easy. But such vectors remain relatively large, have a low copy number and are therefore unsuitable for some cloning purposes. Pdf broadhostrange plasmid vectors for gene expression in. The multi host vector prl83a was used in this study. A new and improved hostindependent plasmid system for rk2. A gene cloning vector should have capability of autonomous replication, small size, origi.
Belongs to a group of vectors atcc 3762237623, 7728777292 differing only in antibiotic resistance and polylinker orientation. Construction of a broad host range cosmid cloning vector. Construction of broadhostrange plasmid vectors for easy visible. Broadhostrange vector pbhr1 mobilizable pbhr1 is a mobilizable vector that was created by removing a frame shift in the mobilization gene of pbbr122, thereby. An autoregulated highlevel expression vector utilizing the tac promoter, rrnb terminators and laciq for lac repression. Broadhostrange plasmid vectors for gene expression in bacteria article pdf available in methods in molecular biology clifton, n.
Potentiation of curing by a broadhostrange selftransmissible. It is derived from rsf1010, a naturally occurring broad host range vector found in e. A versatile onestep crisprcas9 based approach to plasmidcuring. Development of a broadhostrange sacb based vector for. Wed like to understand how you use our websites in order to improve them. Pdf a transposon vector for broad host range cloning. Next generation soil metagenomics largeinsert bac libraries. Concentration of induced gene product may be regulated by varying the lac inducer concentration. Improvement of pbbr1mcs plasmids, a very useful series of. For ease of manipulation and high gene dosage of the cloned material we need small, high copy number, broad host range plasmids. Macrosiphum euphorbiae both species of aphids are attacked by a large range of natural enemies including lady beetles, flower fly larvae, lacewing larvae, predatory midges and parasitic wasps, especially diaeretiella rapae and aphidius nigripes for the green peach and potato aphids, respectively. Cloning vector for copy control fosmid library containing ori2 and oriv, parabc, cos and loxp, cm r, 8.
Four new derivatives of the broad host range cloning vector pbbr1mcs, carrying different antibioticresistance cassettes. Plasmids with rk2 ori region will replicate in most gram negative bacteria plasmids with the rsf 1010 ori can replicate in gram negative and gram. Construction of versatile expression cloning vehicles using the lipoprotein gene of escherichia coli. Broadhostrange vector pbhr1 mobilizable boca scientific. Improvement of pbbr1mcs plasmids, a very useful series of broadhostrange cloning vectors. Development of broadhostrange vectors and gene banks. Rsf 1010 has been completely sequenced scholz 1989. Users may download and print one copy of any publication from the public. Cloning and sequence analysis of homeobox transcription factor. A broad host range cloning vector unknown language. Construction and characterization of a versatile broad host range dna cloning system for gramnegative bacteria. Here we assess a low copy number broadhostrange selftransmissible incp1 plasmid as a vector for such curing cassettes to displace incf. There is an open access version for this licensed article that can be read free of charge and without license restrictions.
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