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Cell Journal، جلد ۱۰، شماره ۳، صفحات ۲۰۱-۲۰۴
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عنوان انگلیسی Cloning Of Leishmania Major P4 Gene
چکیده انگلیسی مقاله Objective: Leishmania major P4 gene is normally expressed during amastigote form of 
the parasite and can be good candidate for producing an effective vaccine. In this study we 
cloned this gene in suitable vector (pQE-30) for further vaccine preparation studies.
Materials and Methods: Leishmania promastigotes were grown in N.N.N.medium and culture in RPMI 1640 cell culture medium. Total genomic DNA was extracted by centrifugation 
of promastigotes. The pellet was suspended in lysis buffer and followed by boiling method. 
PCR was carried out using P4 gene specific primers. PCR product was detected by agaros 
gel electrophoresis and cloned into Bluescript plasmid via T/A cloning method. Reaction 
was transformed into XL1- Blue competent cell and recombinant plasmid screened using 
agar plate contained X-gal and IPTG. The product was extracted, digested by restriction 
enzyme and electrophoresed on agarose gel.
Results: Plasmid was extracted and cloned gene was released by restriction enzyme and 
subcloned into pQE-30 expression vector. 
Conclusion: This construct is ready for protein expression in in-vitro.
کلیدواژه‌های انگلیسی مقاله P4 Gene, Cloning, Leishmania major
نویسندگان مقاله Minoo Shaddel |
Parasitology and Mycology Department, Iran University of Medical Sciences and Health Services, Tehran, Iran

Hormoz Oormazdi |
Parasitology and Mycology Department, Iran University of Medical Sciences and Health Services, Tehran, Iran

Lame Akhlaghi |
Parasitology and Mycology Department, Iran University of Medical Sciences and Health Services, Tehran, Iran

Bahram Kazemi |
Parasitology and Mycology Department, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran

Mojgan Bandehpour |
Molecular and Cellular Biology Research Center, Shahid Beheshti University of Medical Sciences and Health Services, Tehran, Iran

نشانی اینترنتی https://www.celljournal.org/article_248197_6996a3e6587b835fd2fe3fdaa712bbc2.pdf
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