Cloning and expression of recombinant hirudin in CHO cell line

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SHARIFZADEH A.; HEIDARI M.; Aali F.

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Paper Information

Journal: JOURNAL OF COMPARATIVE PATHOBIOLOGY IRAN Year:2020 | Volume:17 | Issue:1 (68) Page(s): 3079-3086

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Title

Cloning and expression of recombinant hirudin in CHO cell line

Author(s)

SHARIFZADEH A. | HEIDARI M. | Aali F. | Issue Writer Certificate

Keywords

HirudnQ1

CloningQ1

gene expressionQ1

Abstract

Hirudin is a 65-66 amino acids polypeptide which is secreted as an anticoagulant compound from salivary glands of medical leech. This drug is a very potent inhibitor of thrombin and is so effective for arterial and venous thrombosis prevention. The aim of the present research was to clone and express of hirudin gene in CHO cell line as a eukaryotic host cell. In this experimental study, a 221 bp fragment of hirudin gene was cloned into pcDNA3. 1(+) vector. The pcDNA3. 1(+)-hirudin recombinant vector was transfected into CHO cells by lipofectamine 2000 reagent. The eukaryotic expression of hirudin was evaluated by RT-PCR method. The pcDNA3. 1(+)-hirudin recombinant vector was confirmed by PCR and enzymatic double digestion. Our findings showed that the mammalian expression of hirudin was successful. As, a 221 bp fragment corresponded to hirudin mRNA was observed on agarose gel after RT-PCR. The pcDNA3. 1(+)-hirudin recombinant vector was constructed in this research successfully. This new recombinant plasmid can express the hirudin mRNA in CHO cells. Therefore, the recombinant protein that produced from this research can be a recombinant vaccine that could be applied for future research. Additionally, this construct has the potential to be use as DNA vaccine in next experiments.

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APA: Copy

SHARIFZADEH, A., HEIDARI, M., & Aali, F.. (2020). Cloning and expression of recombinant hirudin in CHO cell line. JOURNAL OF COMPARATIVE PATHOBIOLOGY IRAN, 17(1 (68) ), 3079-3086. SID. https://sid.ir/paper/389700/en

Vancouver: Copy

SHARIFZADEH A., HEIDARI M., Aali F.. Cloning and expression of recombinant hirudin in CHO cell line. JOURNAL OF COMPARATIVE PATHOBIOLOGY IRAN[Internet]. 2020;17(1 (68) ):3079-3086. Available from: https://sid.ir/paper/389700/en

IEEE: Copy

A. SHARIFZADEH, M. HEIDARI, and F. Aali, “Cloning and expression of recombinant hirudin in CHO cell line,” JOURNAL OF COMPARATIVE PATHOBIOLOGY IRAN, vol. 17, no. 1 (68) , pp. 3079–3086, 2020, [Online]. Available: https://sid.ir/paper/389700/en

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