Design, transformation and proliferation of VEGF111b recombinant isoform in Escherichia coli Top10 in order to produce recombinant drugs

Sadegi, M. and Hojjati, Z. (2016) Design, transformation and proliferation of VEGF111b recombinant isoform in Escherichia coli Top10 in order to produce recombinant drugs. Feyz Journal of Kashan University of Medical Sciences, 20.

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Official URL: http://feyz.kaums.ac.ir/article-1-3179-en.html

Abstract

Background: The synthesis of recombinant proteins with the aim of inhibiting the tumor receptors is one of the new approaches in cancer treatment. The aim of this study was to design and clone the VEGF 111b recombinant isoform in pBudCE4.1 vector and assess its compatibility with E.coli Top10 in order to produce some recombinant drugs. Materials and Methods: The VEGF111b new isoform designed using gene sequences available in the databases and oligo 7 software was digested by BglII and KpnI enzymes. Then it was cloned at downstream of the EF-1 promoter in the pBudCE4.1vector. Isolation of recombinant bacteria was done in LBA medium with the concentration of Zeocin antibiotic (3 and 5). In the final step, the recombinant vector was extracted using DNA gel extraction kit and VEGF111b recombinant fragment was confirmed by enzyme digestion and sequencing. Results: The ligation of 111b fragment in expected site was confirmed. The entire E.coli Top10 colonies were observed in Zeocin medium (5) containing recombinant VEGF111b fragment and recombinant colonies (61.9) were observed at Zeocin medium (3.0). The existence of VEGF111b sequences in recombinant bacteria was confirmed by enzyme digestion and sequencing. Conclusion: The present study is an important step in the production process of VEGF111b recombinant protein and evaluating its anticancer effect. Moreover, the pBudCE4.1 vector containing 2 cloning sites and 8 enzyme excision sites is a good candidate for cloning and expression of recombinant proteins in E.coli TOP10.

Item Type: Article
Subjects: Immunology and Microbiology
Divisions: Feyz journal
Depositing User: ART . editor
Date Deposited: 06 May 2017 15:05
Last Modified: 29 May 2017 15:23
URI: http://eprints.kaums.ac.ir/id/eprint/1455

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