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title

Cloning and Expression of GranzymeM cDNA

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Year: 2014
COI code: CIGS13_0463
Paper Language: English

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Authors Cloning and Expression of GranzymeM cDNA

  Narges Khazeh - Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran
  Mohammad Pazhang - Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran
  Faramarz Mehrnejad - Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran
  Nader Chaparzadeh - Department of Cellular and Molecular Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran

Abstract:

Defence against virally infected and tumor cells depends on the action of cytotoxic T lymphocytes and natural killer cells. GranzymeM is a member of a family of granule serine proteases, which is mainly expressed by these cells. GranzymeM appears to be a potent inducer of tumor cell death with morphological hallmarks that are unique among all granzymes.Since the cloning of the granzymeM cDNA in the early 1990s, it has remained an orphan granzyme for many years and only during the past few years the interest in this protease has increased. Therefore in this study The cDNA of activehGzmM, was cloned into bacterial expression vector pET21a(+).for protein expression, Recombinant plasmid was transformed into E. coli BL21(DE3) cells . Recombinant GranzymeM protein was induced at different conditions. Induction condition was optimized to obtain high yield of recombinant proteinand protein expression analyzed by SDS-PAGE. Results showed that the recombinant enzyme has optimum expression at 22°C with 1mM IPTG during 5h and expressed as inclusion bodies in E. coli

Keywords:

Cloning, Tumor cell, Serine protease, GranzymeM, Inclusion body

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COI code: CIGS13_0463

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Khazeh, Narges; Mohammad Pazhang; Faramarz Mehrnejad & Nader Chaparzadeh, 2014, Cloning and Expression of GranzymeM cDNA, 12th Congress of Iranian Genetics Society, تهران, انجمن ژنتيك ايران, https://www.civilica.com/Paper-CIGS13-CIGS13_0463.htmlInside the text, wherever referred to or an achievement of this article is mentioned, after mentioning the article, inside the parental, the following specifications are written.
First Time: (Khazeh, Narges; Mohammad Pazhang; Faramarz Mehrnejad & Nader Chaparzadeh, 2014)
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Type: state university
Paper No.: 2505
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