STUDY AND EVOLUTION CLONING OF THE GENE ENCODING THE L-ASPARAGINASEΙΙ FROM E. COLI IN B. SUBTILIS

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HOSEINIAN HAMID; BARZAMINY BEHNAZ

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Journal: NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL Year:2014 | Volume:4 | Issue:16 Page(s): 61-66

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Title

STUDY AND EVOLUTION CLONING OF THE GENE ENCODING THE L-ASPARAGINASEΙΙ FROM E. COLI IN B. SUBTILIS

Author(s)

HOSEINIAN HAMID | BARZAMINY BEHNAZ | Issue Writer Certificate

Keywords

L-ASPARAGINASEQ2

E. COLIQ2

B. SUBTILISQ2

CLONINGQ2

PMR12Q1

Abstract

Aim and Background: L-ASPARAGINASEΙΙ (E.C. 3.5.1.1) has effective usage for treatment of acute lymphoblastic leukemia. This enzyme is isolated from bacterial sources and it is commercially available as an anti-cancer drug. This enzyme catalyzes the hydrolysis of L-asparagine to ammonium and aspartate. Unlike Normal cells, the cancer cells strongly require L-Asparagine leading to destruction of these cells. The propose of this project is CLONING of L-ASPARAGINASEΙΙ gene from E. COLI in B. SUBTILIS to produce the mass of this enzyme.Materials and methods: L-ASPARAGINASEΙΙ gene is isolated from E. COLI by PCR approach. The amplified fragment and expression shuttle vector PMR12 are digested by HindΙΙΙ and BamHΙ enzymes. The ligation between DNA fragment and the CLONING shuttle vector is done by standard method. In the next step recombinant vector is transferred to E. COLI JM101 by cold calcium chloride treatment and finally introduced into B. SUBTILIS by a chemical method.Results: in this experiment, ansB gene is isolated from E. COLI by PCR and cloned into the expression shuttle vector PMR12. Existence of gene with 1047 bp lenght is confirmed by enzymatic analysis and PCR reaction. Then recombinant vector cloned in E. COLI at first and then in B. SUBTILIS. Finally the plasmid extracted and compared with the band of expersion shuttle vector containing ansB gene to confirm.Conclusions: In this study, we tried to clone the ansB gene in B. SUBTILIS using expression shuttle vector PMR12. This is the first report of ansB gene CLONING in B. SUBTILIS.

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APA: Copy

HOSEINIAN, HAMID, & BARZAMINY, BEHNAZ. (2014). STUDY AND EVOLUTION CLONING OF THE GENE ENCODING THE L-ASPARAGINASE?? FROM E. COLI IN B. SUBTILIS. NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL, 4(16), 61-66. SID. https://sid.ir/paper/204699/en

Vancouver: Copy

HOSEINIAN HAMID, BARZAMINY BEHNAZ. STUDY AND EVOLUTION CLONING OF THE GENE ENCODING THE L-ASPARAGINASE?? FROM E. COLI IN B. SUBTILIS. NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL[Internet]. 2014;4(16):61-66. Available from: https://sid.ir/paper/204699/en

IEEE: Copy

HAMID HOSEINIAN, and BEHNAZ BARZAMINY, “STUDY AND EVOLUTION CLONING OF THE GENE ENCODING THE L-ASPARAGINASE?? FROM E. COLI IN B. SUBTILIS,” NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL, vol. 4, no. 16, pp. 61–66, 2014, [Online]. Available: https://sid.ir/paper/204699/en

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