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2 edition of Analysis of the streptomyces fertililty plasmid SCP2. found in the catalog.

Analysis of the streptomyces fertililty plasmid SCP2.

Jie Xiao

Analysis of the streptomyces fertililty plasmid SCP2.

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  • 30 Currently reading

Published by University of East Anglia in Norwich .
Written in English


Edition Notes

Thesis(Ph.D.), University of East Anglia, School of Biological Sciences, 1994.

ID Numbers
Open LibraryOL21847168M

  In this work we describe how the detection of an overexpressed RNA by transcriptomic analysis, lead us to the identification of a putative type I TA system in the plasmid DNA of two L. rhamnosus Cited by: 9. To better understand the mechanism of long-palindrome formation in Streptomyces linear plasmids, the pSLA2-derived plasmid pQC26 (Fig. 1; Qin and Cohen ) isolated from S. lividans ZX7 was treated with SacI endonuclease to intentionally produce a site-specific double-strand DNA break between one telomere and the plasmid's replication origin.   Glycopeptide antibiotics inhibit bacterial cell-wall synthesis, and are important for the treatment of infections caused by multi drug-resistant strains of enterococci, streptococci and staphylococci. The main mechanism by which bacteria resist the action of glycopeptides is by producing a modified cell-wall in which the dipeptide D-Alanine-D-Alanine is substituted by D Cited by: 4. Purification and Characterization of a DNA Plasmid Part A Version: Janu INTRODUCTION DNA Plasmids. A plasmid is a small double-stranded, circular DNA molecule that replicates independently of chromosomal DNA. A plasmid must contain an origin of replication, and may Analysis of the agarose gel via restriction mapping will be.


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Analysis of the streptomyces fertililty plasmid SCP2. by Jie Xiao Download PDF EPUB FB2

Gene. 35 () Eisevier GENE The Streptomyces plasmid SCP2 *: its functional analysis and development into useful cloning vectors (Restriction maps; recombinant DNA; hygromycin, thiostrepton and viomycin-resistant derivatives; stable, low-copy-number vectors; E.

coli-Streptomyces shuttle vector) D. Lydiate, F. Malpartida and D.A. Hopwood Cited by: The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors. Lydiate DJ, Malpartida F, Hopwood DA. Detailed restriction maps of the plasmid SCP2* and its deletion derivative pSCP were constructed.

DNA fragments carrying hygromycin (Hyg), thiostrepton (Thio) or viomycin-resistance (VioR) determinants Cited by: The plasmid SCP2, initially discovered through the occurrence of a high fertility variant, SCP2*, is a self-transmissible fertility factor capable of promoting chromosomal recombination within.

Streptomyces lividans ISP contains four small high copy number plasmids: pIJ ( kb), pIJ ( kb), pIJ ( kb) and pIJ ( kb).

The three smaller species appear to be naturally occurring deletion variants of pIJ pIJ and its in vivo and in vitro derivatives were studied after transformation into S. lividans pIJ was found to be self Cited by: Plasmid SCP2* is a 31 kb, circular, low-copy-number plasmid originally identified in Streptomyces coelicolor A3(2) as a fertility factor.

The plasmid was completely sequenced. We have constructed cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp.

All vectors contain the 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 (Am R, Th R, Sp R) that function in Cited by: Streptomyces coelicolor A3(2) possesses two plasmids (SCP1 and SCP2) that act as sex factors.

The plasmid deoxyribonucleic acid isolated from S. coelicolor A3(2) SCP1- strains A and A had the same molecular weight and endonuclease cleavage pattern as the SCP2 plasmid.

The plasmidless strain S18 SCP2- was isolated from the A X A by: 6. Documents recent research and development in streptomycetes genomics, physiology and metabolism.

An excellent source of up-to-date information. Topics include: genome architecture, conjugative genetic elements, differentiation, protein secretion, central carbon metabolic pathways, regulation of nitrogen assimilation, phosphate control of metabolism, gamma.

Lydiate DJ, Malpartida F, Hopwood DA () The Streptomyces plasmid SCP2 *, Analysis of the streptomyces fertililty plasmid SCP2. book functional analysis and development into useful cloning vectors.

Gene – Google Scholar Muth G, Wohlleben W, Pühler A (a) The minimal replicon of the Streptomyces ghanaensis plasmid pSG5 identified by subcloning and Tn5 by: Streptomyces pheochromogenus Streptomyces phaeochromogenes is a bacterium species from the genus of Streptomyces.

[1] [3] [4] Streptomyces phaeochromogenes produces tyrosinate, bromoperoxidase, ditryptophenalin, phaeochromycin A, phaeochromycin B, phaeochromycin C, phaeochromycin D and phaeochromycin : Streptomycetaceae.

The procedure has been used successfully for isolation of plasmids SCP2 and SCP2* as well as the plasmids listed in Table 1 (see next page) from S. Analysis of the streptomyces fertililty plasmid SCP2. book and S. lividans strains.

Genetic manipulation of Streptomyces: a laboratory manual. Hopwood. John Innes Foundation, - Genetic engineering - pages. 0 Reviews. From inside the book. What people medium MgCl2 microcentrifuge mutations mycelium NaCl nitrocellulose nucleic acid Pasteur pipette pellet phage phenol plaques plasmid plates pocks precipitation.

It makes the analysis of future reactions easier, for example, the viewing of small fragments after a restriction digest. BioCoder version Following is the Small Scale Plasmid Isolation (Mini-Maxi prep) protocol in BioCoder, a high-level programming language for expressing biology protocols.

Abstract. The methyl-specific restriction system of Streptomyces coelicolor A3(2) was analyzed by carrying out transformations with unmethylated and methylated pSET omyces coelicolor was found to strongly restrict DNA methylated in vivo by the Dam, Dcm and Hsd modification systems of Escherichia -modified DNA was restricted as Cited by:   Plasmid pNGem3-Anti, a multicopy Streptomyces plasmid that harbours the yefMsl gene under the control of the xylanase promoter xysAp (see Methods), was used to make the expression plasmids by cloning the genes encoding the Amy α-amylase or the Xys1 xylanase under the control of the pstSp promoter (see Methods).

Plasmid Curing from Streptomyces sp: The removal of plasmid from bacterial cells lead to missing of certain characters of these cells such as antibiotic resistance or antibiotic production.

In the present study the treatment of Streptomyces sp. with acridine orange (28 g /ml) result in. Integrating vectors based on the int/attP loci of temperate phages are convenient and used widely, particularly for cloning genes in Streptomyces spp.

We have constructed and tested a novel integrating vector based on g27, encoding integrase, and attP site from the phage, SV1. This plasmid, pBF3 integrates efficiently in S. coelicolor and S. lividans but surprisingly Cited by: Journal of' General Microbiology (),Printed in Great Britain Temperature-sensitive Mutants of the Streptomyces Plasmid pIJ By ASHLEY W.

BIRCH AND JOHN CULLUM* Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute. The plasmid contains the nat1 gene from Streptomyces noursei encoding nourseothricin N-acetyl-transferase and confers resistance to the antibiotic nourseothricin of transformed yeasts.

Other plasmids conferring nourseothricin resistance: pAG35, pAG Plasmid usefull as template for PCR to create gene deletions. Plasmid SCP2-pUC from Dr. James Kadonaga's lab contains the insert SCP2 (super core promoter 2)-pUC and is published in Nat Methods. Nov. 3(11) This plasmid is available through Addgene.

Sequencing and analysis of pWTY27 The unique SacI-treated pWTY27 was cloned in an E. coli plasmid pSP72 for shotgun cloning and sequencing (see Methods). The complete nucleotide sequence of pWTY27 consisted of 14, bp with % GC content, resem-bling that of a typical Streptomyces genome (e.g.

% for S. coelicolor) [25]. 2 Protocol 1. Strains and plasmids for recombineering in Streptomyces coelicolor Strain/ Plasmid Relevant Genotype/Comments1 Source/ Reference Plasmids pIJ λ-RED (gam, bet, exo), cat, araC, repts Gust et al., pIJ P1-FRT-oriT-aac(3)IV-FRT-P2 Gust et al., pIJ P1-FRT-oriT-neo-FRT-P2 Gust et al., pIJ P1-FRT-neo-FRT-P2 Gust et al., File Size: KB.

Practical Streptomyces Genetics Paperback – January 1, See all formats and editions Hide other formats and editions. Price New from Used from Paperback, January 1, Format: Paperback.

prehensive mutational analysis to confirm the identity and function of the SRB biosynthetic genes is in progress in our laboratory.

Fig. (A) Chemical structure of lankacidin C (LC) and lankamycin (LM) isolated from Streptomyces rochei AN4.

(B) Gene organization of. A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 20% sterile glycerol at °C. It will last for many years if it is prepared in a sterile environment and afterwards, treated carefully.

ing. Conjugation in Streptomyces produces pocks which are zones of transient inhibition of growth of recipients. Kieser et al. () local~ ized the transfer and spread regions, involved in plasmid transfer and pock formation.

respective­ ly, in plJa multi-copy, broad host range plasmid from Streptomyces fividallTJ). Subse­ quently. Plasmids A Desktop Resource (1st Edition) 2 | P a g e Plasmids Introduction to Addgene’s Resource Any newcomer who joins a molecular biology lab will undoubtedly be asked to design, modify, or construct a plasmid.

Although the newcomer likely knows that a plasmid is a small circular piece of DNA found in bacterial cells, she mayFile Size: 2MB. Do you have a protocol for the isolation of very low-copy plasmids from Streptomyces spp.

FAQ ID Yes, please follow the User-Developed Protocol ' Isolation of very low-copy plasmids from Streptomyces spp. using the QIAGEN Plasmid Maxi Kit ' (QP13). The strategy for PCR-targeting for mutagenesis of Streptomyces coelicolor.

is to replace a chromosomal sequence within a. coelicolor. cosmid (Redenbach. et al., ) by a selectable marker that has been generated by PCR using primers with 39 nt homology extensions.

The inclusion of. oriT (RK2) in the disruption cassette allowsFile Size: KB. Four different extraction methods of small plasmid DNA from antibiotic-producing Streptomyces isolates and from the positive control S.

lividans, containing the pIJ plasmid, were standardized. Among these, only one procedure allowed the detection of plasmid DNA from the positive control very effectively that was the Kieser () method. BHATTACHARYYA & SEN: GENE MANIPULATION IN STREPTOMYCES 23 al45 constructed a recombinant plasmid pNL The construction was done with a kb fragment of S.

ansochromogenes, which was involved in nikkomycin biosynthesis and. plasmid el host cell transformed host streptomyces Prior art date Legal status (The legal status is an assumption and is not a legal conclusion.

Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) Expired Application number CAA Other languages French (fr) InventorCited by: Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E.

coli ET (pUZ), as a conjugal donor, carrying the integrative plasmid pSET For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces Cited by: 9. Plasmid replication is a rigorously controlled process in part because plasmid overreplication would tax the metabolic capacity of the host cell and put the plasmid-bearing cell at a disadvantage compared to a plasmid-free counterpart.

Plasmids con-trol their copy number primarily at the stage of replication initiation. The frequencyFile Size: 3MB.

DNA purification and isolation of genomic DNA from DNA analysis by multiplex and real-time PCR, the importance of high-quality, purified DNA cannot be underestimated.

Finding a suitable DNA isolation system to satisfy your downstream application needs is vital for plasmid DNA contained in the cleared alkaline lysates. plasmid host cell pel7 streptomyces recombinant dna Prior art date Legal status (The legal status is an assumption and is not a legal conclusion.

Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) Expired - Fee Related Application number US06/, Inventor Walter M Cited by: OVER 60% of known antibiotics1 are produced by Streptomyces species, including many substances with valuable clinical and other applications; they therefore have considerable medical, biological.

Genetic Manipulation of Streptomyces: A Laboratory Manual Plastic Comb – June 1, by D. Hopwood (Author), M. Bibb (Author), K. Chater (Author), & See all formats and editions Hide other formats Author: D. Hopwood, M. Bibb, K. Chater.

Characterization of the minimal origin required for replication of the streptococcai plasmid plP in Bacillus subtilis S. Brantl^ and D. Behnke Institute for Moiecuiar Biology, Beutenbergstr Jena, Germany.

Summary By using deletional analysis the origin of replication, oriR, of the streptococcai plasmid plP in Bacillus. The genetic information needed for a cell to participate in conjugation resides in the DNA of a cell's _____.

ondria b.F plasmid c.F pilus. TECHNIQUES IN MOLECULAR BIOLOGY – METHODS FOR PLASMID DNA ISOLATION 3 Other notes on this plasmid mini-prep technique – Once cells have been lysed, mixing should be done thoroughly but gently, to avoid breaking plasmid and bacterial chromosomal DNA.

Do not vortex after cell resuspension, but mix by inversion. – After the protein precipitation step, the File Size: KB.Vector IG Sequence Link: General: plasmid ds-DNA BP Functions: (cloning) Selection: () Copy Number: Hosts: (Streptomyces lividans )(Streptomyces lividans) Suppliers: (ATCC) ts: The tyrosinase (mel) gene of S.

antibioticus was inserted at a BclI site of pIJ, a non-conjugative broad host range vector. Tyrosinase is secreted during growth of S. Novel chemical compounds, recombinant plasmids pUC and pUC, which are obtained by covalent linkage of the E.

coli plasmid pBR to the Streptomyces espinosus plasmid pUC6. These plasmids are produced by a novel process which can be used to stabilize unstable potential plasmid vectors.