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

Publish Year: 1398
نوع سند: مقاله کنفرانسی
زبان: English
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DCME02_270

تاریخ نمایه سازی: 19 آذر 1398

Abstract:

Background:Malaria is a vector borne infectious disease throughout the world that affects the human and animal. Plasmodiumspp. and Anopheles spp. are the two key elements in malaria emergence which are the cause and vector of the disease respectively. These items are the target of control strategies, to disrupt the parasite life cycle or transmission. Concerning the broad social and economic impacts of malaria, second global malaria eradication program was managed by World Health Organization (WHO). To reach this goal, different strategies were recommended by scientists. Development of vaccines that interrupt malaria transmission (VIMTs) was one of them. VIMTs are divided to different groups that the vector based vaccines are a member of them. According to the important role of Anopheles carboxypeptidase in sexual parasite development and the role of Anopheles stephensi in malaria transmission, we characterized the first member of cpb gene family (cpbAs1) of An. stephensi to provide the basic information for development an effective vector based VIMT.Objective: Cloning, expression and characterization of thecpbAs1of An. Stephensito provide the basic information for development an effective vector based VIMT.Materials and methods:At the first, the middle part sequence of cpbAs2 mRNA sequence was determined. Then, its 3 - and 5 -ends were sequenced by 3 - and 5 -RACE methods respectively. Next, sequence have been amplifiedwith 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 carboxypeptidase 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.

Keywords:

Malaria , Anopheles , Carboxypeptidase , Vector control , Vaccines that Interrupt Malaria Transmission

Authors

Zahra Ghorbanzadeh

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

Maryam derikvand

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