CLONING AND EXPRESSION OF CARBOXYPEPTIDASE B1 OF ANOPHELES STEPHENSI THE MAIN MALARIA VECTOR IN IRAN

سال انتشار: 1398
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 451

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شناسه ملی سند علمی:

DCME02_060

تاریخ نمایه سازی: 19 آذر 1398

چکیده مقاله:

Background: Malaria is the most important protozoan infectious disease in human and World Health Organization have some objectives towards the elimination of Malaria till 2050. One of the main object is use the VIMT vaccines. Carboxypeptidase Enzymes have main role in digestion of proteins in the midgut of the Anopheles and interruption in the normal function of this enzymes may cause the significance effect on the plasmodium life cycle and anopheles fitness. It is expected that with the inhibition of simultaneous activity of both cb1&cb2 enzymes, the inhibition of their activity could be synergically increased. At this investigation we clone and express the potato carboxypeptidase inhibitor (PCI) to produce large scale for inhibition assay of enzymes.Objective: cloning and expression of the potato carboxypeptidase inhibitor (PCI) for inhibition assay of Carboxypeptidase cb1 and cb2.Materials and methods: At the first, the sequences reported for PCI were extracted from Genbank and aligned. Then, bioinformatics analysis were done and the coding region will be sent to gene synthesis. The sequences have been amplified with PCR, T/A cloned and ligated and then transferred to PET-28 expression vector to express as a recombinant protein in E.coli.Verification of recombinant protein expression was performed by SDS-page and Western Blotting techniques.Results and Discussion: The potato carboxypeptidase inhibitor gene was successfully amplified by PCR and then cloned into the pET-28a expression vector. The expression of recombinant protein expression was confirmed by SDS-PAGE and Western blotting assays.Conclusion: As mentioned before, we can make suitable effects on the life cycle of the plasmodium parasites in the midgut of anopheles by using specific inhibitor against one of the main peptidase family. Also, we can make some changes in the life cycle and fitness of the Anopheles that can help us to decrease the malaria cases in the endemic areas.

کلیدواژه ها:

Malaria ، potato carboxypeptidase inhibitor (PCI) ، Anopheles ، Carboxypeptidase ، Vector control

نویسندگان

Maryam Drikvand

Laboratory science BSc, Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran, Iran

Abbasali Raz

Associated professor, Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran, Iran

Mahdokt Ilbeigikhamsenejad

Cellular and Molecular biology MSCs , Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran, Iran.

Navid Dinparast D.jadid

Professor, Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran, Iran

Zahra Ghorbanzadeh

Laboratory science BSc, Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur Institute of Iran, Tehran, Iran.