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Friday, July 24, 2020 | History

2 edition of Cloning and expression of foreign genes in mycobacteria found in the catalog.

Cloning and expression of foreign genes in mycobacteria

Odir Antonio Dellagostin

Cloning and expression of foreign genes in mycobacteria

by Odir Antonio Dellagostin

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Published by University of Surrey in Guildford .
Written in English


Edition Notes

Thesis (Ph.D) - University of Surrey,1994.

StatementOdir Antonio Dellagostin.
ContributionsUniversity of Surrey.
ID Numbers
Open LibraryOL19593018M

  Recombineering in Mycobacterium tuberculosis. nonessential M. tuberculosis groEL1 gene (c) was generated by cloning Gene replacement and expression of foreign DNA in mycobacteria. The cloning and functional expression of Mycobacterium tuberculosis Rvc in Escherichia coli revealed that this gene encodes the diterpene cyclase for producing (+)-5(6),halimadieneol, which accepts geranylgeranyldiphosphate as the intrinsic substrate.

Gene replacement by homologous recombination (HR) is an invaluable tool in understanding the physiology and the significance of specific genes in the virulence of Mycobacterium tuberculosis. It will also allow for the development of rationally attenuated strains as candidate vaccines to prevent the spread of tuberculosis. A 92 kb cryptic Mycobacterium fortuitum plasmid, pMF1, was isolated from strain and its restriction map constructed. A 42 kb HindIII fragment of pMF1 was found to support replication in mycobacteria and this fragment was cloned and sequenced to characterize the replication elements of the plasmid. Computer analysis identified a putative Rep protein ( amino acids) with high homology.

A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria; however, plasmids are sometimes present in archaea and eukaryotic nature, plasmids often carry genes that benefit the survival of the organism. Rivera-Marrero CA, Ritzenthaler JD, Newburn SA, Roman J, Cummings RD. Molecular cloning and expression of a novel glycolipid sulfotransferase in Mycobacterium tuberculosis. Microbiology. ; (Pt 3)


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Cloning and expression of foreign genes in mycobacteria by Odir Antonio Dellagostin Download PDF EPUB FB2

The potential use of the cloned gene in construction of genetically defined mutant strains of M. tuberculosis by gene replacement is proposed as a novel approach to the rational attenuation of mycobacterial pathogens and the possible development of new antimycobacterial by:   This report describes the first successful transfer and complete expression of clustered mycobacterial genes controlling a biosynthetic pathway (carotenogenesis) in a homologous system.

A genomic library of pigmented Mycobacterium aurum A + (yellow-orange) DNA was constructed in shuttle vector pHLDCited by: The methods range from basic culture techniques, cell fractionation, and nucleic acid purification to more advanced protocols for the introduction of cloned DNA into mycobacteria, the use of reporter genes, and the expression of foreign genes.

Asian Pacific Journal of Tropical Medicine () Document heading doi: Cloning, expression, identification and bioinformatics analysis of Rvc gene from Mycobacterium tuberculosis in Escherichia coli Qiang Wu 1,2,3, Peng Zhou 1,2*, Shiyun Qian 3, Xi Qin 4, Zhigang Fan Cloning and expression of foreign genes in mycobacteria book, Qiongyao Fu 3, Zhinong Zhan 3, Hua Pei 3 1 Agriculture School of Hainan University, Haikou Cited by: 7.

Cloning of these genes has been achieved by functional complementation. However, clon- ing of mycobacterial genes by functional complementa- tion of well characterized Escherichia coli (Ec) mutant strains has met with limited success, presumably due to poor mycobacterial gene expression in Ec (Clark-Curtiss et al., ).Cited by: Abstract.

Over the past 10 years, major advances have been made in the field of molecular mycobacterioiogy. These include the generation of shuttle vectors for use in both Escherichia coli and mycobacteria, electroporation protocols to introduce these plasmids into mycobacteria and promoter cassettes, which can be used to drive the expression of cloned genes in the mycobacterial host (1.

The study of mycobacterial genomes has exploded during the last 10 yr. Initially, no systems were available for the du-ect manipulation of mycobacterial genes in mycobactena, so Escherichia coliwas. Therefore there is an increased need for highly efficient methods for cloning of mycobacterial genes, as the limited cloning flexibility of current Escherichia coli–mycobacteria shuttle vectors remains a frequent impediment in genetic manipulation of mycobacteria.

Here the authors present a CRISPR interference approach that is able to efficiently repress the expression of target genes in mycobacteria, in a rapid and cost-effective manner.

By identifying an herbicide resistance gene and cloning it into a plant expression vector system, like the Ti plasmid system from Agro bacteriumtume faciens. The scientist would then introduce it into the plant cells by transformation, and select cells that have taken up and integrated the herbicide-resistance gene into the genome.

FEMS Microbiology Letters 30 () Published by Elsevier FEM Cloning and expression of mycobacterial plasmid DNA in Escherichia coli (Cloning; expression; Mycobacterium fortuitum; plasmid DNA) A.

Labidi, H.L. David, and D. Roulland-Dussoix Service de la Tuberculose et des Mycobacties, and * Groupement du Gie Gique, Institut Pasteur, 25 Rue du Dr. Roux, ?4 Paris. Curcic, R., Dhandayuthapani, S, and Deretic, V () Gene expression in mycobacteria. transcriptional fusions based on xyIE and analysts of the promoter region of the response regulator mtrA from Mycobacterium tuberculosis Mol Microb – Google Scholar.

A system that permits molecular genetic manipulation of mycobacteria was developed on the basis of the yeast paradigm of gene replacement by homologous recombination. A shuttle vector that can replicate autonomously at a high copy number in Escherichia coli but must integrate into homologous DNA for survival in Mycobacterium smegmatis was by: The gene for the extracellular alpha antigen of Mycobacterium bovis BCG was cloned by using a single probe restricted to G or C in the third position.

This technique should have great potential for the isolation of mycobacterial antigen by: M. bovisBCG is a strong immunostimulant and has been used as an adjuvant in various protocols of immunization (23).

The recent development of genetic tools for mycobacteria has enabled the cloning of foreign genes in fast-growing mycobacteria such as Mycobacterium smegmatismc and in Cited by: 2. The first is the inability to regulate high-level expression of foreign genes in M.

smegmatis, analogous to systems such as induction of the lac promoter in E. coli. Secondly, no simple, efficient and widely adaptable method for the purification of proteins from recombinant mycobacteria has been described. Systematic and Applied Microbiology 30 () – Mycobacterium species identification – A new approach via dnaJ gene sequencing$ Makiko Yamada-Nodaa,b, Kiyofumi Ohkusua, Hiroyuki Hatac, Mohammad Monir Shaha, Pham Hong Nhunga, Xiao Song Suna, Masahiro Hayashia, Takayuki Ezakia, aDepartment of Microbiology, Regeneration and Advanced Medical Science, Gifu University Graduate.

Intracellular survival plays a central role in the pathogenesis of Mycobacterium tuberculosis. To identify M. tuberculosis genes required for intracellular survival within macrophages, an M.

tuberculosis H37Rv plasmid library was constructed by using the shuttle vector pOLYG. This plasmid library was electroporated into Mycobacterium smegmatis c, and the transformants were used to infect. Abstract.

Despite many methodological advances that have facilitated investigation of Mycobacterium tuberculosis pathogenesis, analysis of essential gene function in this slow-growing pathogen remains difficult.

Here, we describe an optimized CRISPR-based method to inhibit expression of essential genes based on the inducible expression of an enzymatically inactive Cas9 protein together with.

The regions of this gene encoding an amino-acid sequence corresponding to mature TNF have been expressed in Escherichia coli and the product of this expression isolated in pure form and shown to. A Riboswitch-Based Inducible Gene Expression System for Mycobacteria Article (PDF Available) in PLoS ONE 7(1):e January with Reads How we measure 'reads'.Its gene was cloned by using a previously developed single-probe method and was sequenced.

The gene encoded a amino-acid polypeptide with a molecular weight of 21, A recombinant Avi-3 antigen expressed in Escherichia coli reacted with monoclonal and polyclonal antibodies raised against the native Avi-3 by: Thereafter, the expression of the lacZ reporter gene in M.

vaccae, Mycobacterium bovis BCG and Mycobacterium smegmatis mc(2) was evaluated using a .