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Kenneth M. Peterson - One of the best experts on this subject based on the ideXlab platform.

  • Four new derivatives of the broad-host-range Cloning Vector pBBR1MCS, carrying different antibiotic-resistance cassettes
    Gene, 1995
    Co-Authors: Michael E. Kovach, D. Steven Hill, Michael A. Farris, R. Martin Roop, Philip H. Elzer, Gregory T Robertson, Kenneth M. Peterson
    Abstract:

    Four new antibiotic-resistant derivatives of the broad-host-range (bhr) Cloning Vector pBBR1MCS have been constructed. These new plasmids have several advantages over many of the currently available bhr Vectors in that: (i) they are relatively small (< 5.3 kb), (ii) they possess an extended multiple Cloning site (MCS), (iii) they allow direct selection of recombinant plasmid molecules in Escherichia coli via disruption of the LacZα peptide, (iv) they are mobilizable when the RK2 transfer functions are provided in trans and (v) they are compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1- and P15a-based replicons. © 1995.

  • In vivo and in vitro stability of the broad-host-range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, R. Martin Roop
    Abstract:

    Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus. To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella. pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms. © 1995 Academic Press, Inc.

  • in vivo and in vitro stability of the broad host range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, Martin R Roop
    Abstract:

    Abstract Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus . To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella . pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms.

Michael E. Kovach - One of the best experts on this subject based on the ideXlab platform.

  • four new derivatives of the broad host range Cloning Vector pbbr1mcs carrying different antibiotic resistance cassettes
    Genes, 1995
    Co-Authors: Michael E. Kovach, Philip H. Elzer, Erin M Farris, D Hill, Philip G Robertson, Christy J Peterson, Dennis R Roop
    Abstract:

    Four new antibiotic-resistant derivatives of the broad-host-range (bhr) Cloning Vector pBBRIMCS have been constructed. These new plasmids have several advantages over many of the currently available bhr Vectors in that: (i) they are relatively small (< 5.3 kb), (ii) they possess an extended multiple Cloning site (MCS), (iii) they allow direct selection of recombinant plasmid molecules in Escherichia eoli via disruption of the LacZct peptide, (iv) they are mobilizable when the RK2 transfer functions are provided in trans and (v) they are compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1and P15a-based replicons.

  • Four new derivatives of the broad-host-range Cloning Vector pBBR1MCS, carrying different antibiotic-resistance cassettes
    Gene, 1995
    Co-Authors: Michael E. Kovach, D. Steven Hill, Michael A. Farris, R. Martin Roop, Philip H. Elzer, Gregory T Robertson, Kenneth M. Peterson
    Abstract:

    Four new antibiotic-resistant derivatives of the broad-host-range (bhr) Cloning Vector pBBR1MCS have been constructed. These new plasmids have several advantages over many of the currently available bhr Vectors in that: (i) they are relatively small (< 5.3 kb), (ii) they possess an extended multiple Cloning site (MCS), (iii) they allow direct selection of recombinant plasmid molecules in Escherichia coli via disruption of the LacZα peptide, (iv) they are mobilizable when the RK2 transfer functions are provided in trans and (v) they are compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1- and P15a-based replicons. © 1995.

  • In vivo and in vitro stability of the broad-host-range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, R. Martin Roop
    Abstract:

    Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus. To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella. pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms. © 1995 Academic Press, Inc.

  • in vivo and in vitro stability of the broad host range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, Martin R Roop
    Abstract:

    Abstract Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus . To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella . pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms.

Philip H. Elzer - One of the best experts on this subject based on the ideXlab platform.

  • four new derivatives of the broad host range Cloning Vector pbbr1mcs carrying different antibiotic resistance cassettes
    Genes, 1995
    Co-Authors: Michael E. Kovach, Philip H. Elzer, Erin M Farris, D Hill, Philip G Robertson, Christy J Peterson, Dennis R Roop
    Abstract:

    Four new antibiotic-resistant derivatives of the broad-host-range (bhr) Cloning Vector pBBRIMCS have been constructed. These new plasmids have several advantages over many of the currently available bhr Vectors in that: (i) they are relatively small (< 5.3 kb), (ii) they possess an extended multiple Cloning site (MCS), (iii) they allow direct selection of recombinant plasmid molecules in Escherichia eoli via disruption of the LacZct peptide, (iv) they are mobilizable when the RK2 transfer functions are provided in trans and (v) they are compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1and P15a-based replicons.

  • Four new derivatives of the broad-host-range Cloning Vector pBBR1MCS, carrying different antibiotic-resistance cassettes
    Gene, 1995
    Co-Authors: Michael E. Kovach, D. Steven Hill, Michael A. Farris, R. Martin Roop, Philip H. Elzer, Gregory T Robertson, Kenneth M. Peterson
    Abstract:

    Four new antibiotic-resistant derivatives of the broad-host-range (bhr) Cloning Vector pBBR1MCS have been constructed. These new plasmids have several advantages over many of the currently available bhr Vectors in that: (i) they are relatively small (< 5.3 kb), (ii) they possess an extended multiple Cloning site (MCS), (iii) they allow direct selection of recombinant plasmid molecules in Escherichia coli via disruption of the LacZα peptide, (iv) they are mobilizable when the RK2 transfer functions are provided in trans and (v) they are compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1- and P15a-based replicons. © 1995.

  • In vivo and in vitro stability of the broad-host-range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, R. Martin Roop
    Abstract:

    Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus. To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella. pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms. © 1995 Academic Press, Inc.

  • in vivo and in vitro stability of the broad host range Cloning Vector pbbr1mcs in six brucella species
    Plasmid, 1995
    Co-Authors: Philip H. Elzer, Michael E. Kovach, Robert W. Phillips, Kenneth M. Peterson, Gregory T Robertson, Martin R Roop
    Abstract:

    Abstract Previous studies have shown that the broad-host-range plasmid pBBR1MCS can be used for genetic complementation in Brucella abortus . To extend these observations, the in vivo and in vitro stability of pBBR1MCS was evaluated in the six currently recognized species of the genus Brucella . pBBR1MCS was readily introduced into all of the strains tested by electroporation and was stably maintained in broth cultures without antibiotic selection during five serial passages over a 10-day period. Furthermore, isolates of all six Brucella strains containing pBBR1MCS obtained from the spleens of BALB/c mice 1 week postinfection maintained the plasmid. Although pBBR1MCS maintains the mobilization locus present in the parental plasmid pBBR1CM, attempts to detect transfer of pBBR1MCS between Brucella strains by conjugation were unsuccessful. These results demonstrate the in vitro and in vivo stability of pBBR1MCS in Brucella spp. and reinforce the usefulness of this Cloning Vector for the genetic analysis of these organisms.

Zahra Meshkat - One of the best experts on this subject based on the ideXlab platform.

  • Design and Construction of a Eukaryotic Cloning Vector Encoding the mpt51 Gene of Mycobacterium tuberculosis.
    Reports of biochemistry & molecular biology, 2019
    Co-Authors: Faria Hasanzadeh Haghighi, Hosna Zare, Ehsan Aryan, Aida Gholoobi, Zahra Meshkat
    Abstract:

    Background: Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a Cloning Vector encoding Mycobacterium tuberculosis (MTB) mpt51 gene. Methods: DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the mpt51 gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) Cloning Vector were both digested with restriction enzymes, the mpt51 fragment was ligated into the Vector, and the Escherichia coli (E. coli) TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing. Results: The mpt51 gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/mpt51 recombinant plasmid and a 926 bp band for mpt51 were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the mpt51 fragment of H37Rv in GenBank. Conclusion: In the current study, the mpt51 gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant Vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this Vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.

  • Design and Construction of a Cloning Vector Containing the hspX Gene of Mycobacterium tuberculosis.
    Reports of biochemistry & molecular biology, 2016
    Co-Authors: Atieh Yaghoubi, Ehsan Aryan, Hosna Zare, Shadi Alami, Roghayeh Teimourpour, Zahra Meshkat
    Abstract:

    BACKGROUND: Tuberculosis (TB) is a major cause of death worldwide. Finding an effective vaccine against TB is the best way to control it. Several vaccines against this disease have been developed but none are completely protective. The aim of this study was to design and construct a Cloning Vector containing the Mycobacterium tuberculosis (M. tuberculosis) heat shock protein X (hspX). METHODS: First, an hspX fragment was amplified by PCR and cloned into plasmid pcDNA3.1(+) and recombinant Vector was confirmed. RESULTS: A 435 bp hspX fragment was isolated. The fragment was 100% homologous with hspX of M. tuberculosis strain H37Rv in GenBank. CONCLUSION: In this study, the Cloning Vector pcDNA3.1(+), containing a 435-bp hspX fragment of M. tuberculosis, was constructed. This could be used as a DNA vaccine to induce immune responses in animal models in future studies.

  • Determination of human papillomavirus type 16 genotype and construction of Cloning Vector pTZ57R encoding HPV16 E7 gene.
    Saudi Medical Journal, 2007
    Co-Authors: Zahra Meshkat, Hessam Mirshahabi, Zuhair M. Hassan, Saeed R. Ghaffari, Mahmoud Mahmoudi, Hoorieh Soleimanjahi, Tayebeh Sabokbar
    Abstract:

    OBJECTIVES To isolate and construct a Cloning Vector containing the human papillomavirus (HPV)16-E7 gene as a target for application as a DNA vaccine. METHODS The study was performed in 2005 in Iran. The E7 gene, one of the most important HPV oncoproteins and a target molecule for therapeutic vaccines, was amplified by polymerase chain reaction (PCR). The PCR product was cloned into a suitable Cloning Vector and confirmed by colony-PCR, restriction enzyme analysis, and sequenced. RESULTS The desired plasmid was sequenced and indicated 99% homology with those mentioned in the Genbank. CONCLUSIONS The Iranian HPV16 E7 gene sequence is very similar to other sequences in the Genbank, and it can be used as a candidate gene in a therapeutic vaccine for Iranian patients with cervical cancer.

  • Determination of human papillomavirus type 16 genotype and construction of Cloning Vector pTZ57R encoding HPV16 E7 gene
    Saudi Medical Journal, 2007
    Co-Authors: Zahra Meshkat, Hessam Mirshahabi, Zuhair M. Hassan, Saeed R. Ghaffari, Mahmoud Mahmoudi, Hoorieh Soleimanjahi, Tayebeh Sabokbar
    Abstract:

    Objective: To isolate and construct a Cloning Vector containing the human papillomavirus (HPV)16-E7 gene as a target for application as a DNA vaccine. Methods: The study was performed in 2005 in Iran. The E7 gene, one of the most important HPV oncoproteins and a target molecule for therapeutic vaccines, was amplified by polymerase chain reaction (PCR). The PCR product was cloned into a suitable Cloning Vector and confirmed by colony-PCR, restriction enzyme analysis, and sequenced. Results: The desired plasmid was sequenced and indicated 99% homology with those mentioned in the Genbank. Conclusion: The Iranian HPV16 E7 gene sequence is very similar to other sequences in the Genbank, and it can be used as a candidate gene in a therapeutic vaccine for Iranian patients with cervical cancer.

Isabelle Derré - One of the best experts on this subject based on the ideXlab platform.

  • A C. trachomatis Cloning Vector and the Generation of C. trachomatis Strains Expressing Fluorescent Proteins under the Control of a C. trachomatis Promoter
    PLoS ONE, 2013
    Co-Authors: Hervé Agaisse, Isabelle Derré
    Abstract:

    Here we describe a versatile Cloning Vector for conducting genetic experiments in C. trachomatis. We successfully expressed various fluorescent proteins (i.e. GFP, mCherry and CFP) from C. trachomatis regulatory elements (i.e. the promoter and terminator of the incDEFG operon) and showed that the transformed strains produced wild type amounts of infectious particles and recapitulated major features of the C. trachomatis developmental cycle. C. trachomatis strains expressing fluorescent proteins are valuable tools for studying the C. trachomatis developmental cycle. For instance, we show the feasibility of investigating the dynamics of inclusion fusion and interaction with host proteins and organelles by time-lapse video microscopy.