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:: Volume 20, Issue 5 (Bimonthly 2016) ::
Feyz Med Sci J 2016, 20(5): 447-453 Back to browse issues page
Design, transformation and proliferation of VEGF111b recombinant isoform in Escherichia coli Top10 in order to produce recombinant drugs
Morteza Sadegi * , Zohreh Hojjati
Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, I. R. Iran. , ms.sadeghi@yahoo.com
Abstract:   (4178 Views)

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.

Keywords: E.coli, Recombinant Protein, VEGF111b
Full-Text [PDF 581 kb]   (2872 Downloads)    
Type of Study: Research | Subject: medicine, paraclinic
Received: 2016/11/13 | Revised: 2017/07/29 | Accepted: 2016/11/13 | Published: 2016/11/13
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Sadegi M, Hojjati Z. Design, transformation and proliferation of VEGF111b recombinant isoform in Escherichia coli Top10 in order to produce recombinant drugs. Feyz Med Sci J 2016; 20 (5) :447-453
URL: http://feyz.kaums.ac.ir/article-1-3179-en.html


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Volume 20, Issue 5 (Bimonthly 2016) Back to browse issues page
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