Molecular Cloning and Expression of the Leishmania infantum KMP-11 Gene

Soflaei, Saied; Dalimi, Abdolhossein; Ghafarifar, Fatemeh

doi:10.5812/jjm.4798

Cited by 2 publications

(2 citation statements)

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“…The cell were lysed by sonication 30", 60 pals and centrifuged at 16000 g for 20 min at 4°C to separate the insoluble proteins from the soluble ones. Finally, the supernatant and pellet were analyzed by SDS-PAGE [2].…”

Section: Expression Of Recombinant Proteinmentioning

confidence: 99%

“…Humans, domestic and wild animals could be infected by VL and found throughout parts of the old and new worlds [1]. VL is characterized by the presence of; fever, hepatosplenomegaly, swollen lymph nodes and weight loss, depending on the Leishmania species and the host's immune response against the parasite [2]. Different methods are used for the diagnosis of VL based on aspirates or biopsy specimens of visceral tissues (spleen, liver, bone marrow), which are subjected to microscopic examination and culture, and serological methods such as the indirect fluorescent antibody (IFA), the indirect hemagglutination assay (IHA), the direct agglutination test (DAT) and enzyme-linked immunosorbent assay (ELISA) [3].…”

Section: Introductionmentioning

confidence: 99%

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Cloning, expression and purification of recombinant A2 protein from Leishmania infantum for diagnosis of visceral leishmaniasis in Iran

Naderi¹,

Nahrevanian²,

Khalaj³

et al. 2016

asb

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Visceral leishmaniasis (VL) is a fatal disease caused by Leishmania infantum in the Mediterranean basin and Iran. Different methods are used for diagnosis of VL. The aims of this study were expression and purification of recombinant A2 (rA2) protein of L.infantum and its application in the diagnosis of VL. The serological diagnosis of VL was applied using rA2 protein. In this study, A2 gene of L.infantum sequence was ordered for the synthesis, cloned in E.coli strain TOP10F' and proliferated in pET22-b vector. The expression and purification of rA2 proteins applied in host via BL21 and Ni-NTA respectively. The A2 gene sequences were synthesized and the construct transformed to pET22-b vector. A 102 Sanaz Naderi et al. 520bp fragment was identified in digested pEASY-A2 plasmid. The gene was successfully cloned in to pET22-b standard expression vector and transformed in E.coli BL21. Expression of rA2 was confirmed by SDS-PAGE and a 27KD protein was detected. The antigenicity of A2 protein was assessed using both pooled dog sera and C9 anti-A2 monoclonal Ab. This study recommends rA2-ELISA as alternative assay to detect VL. More evaluation should be made to develop a cheap and reliable serologic test for detection of L.infantum among infected hosts.

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Section: Expression Of Recombinant Proteinmentioning

confidence: 99%