Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Recombinant plasmid KMP-11 gene of Leishmania major (pcKMP-11): production, characterization and sequencing

(2014) Recombinant plasmid KMP-11 gene of Leishmania major (pcKMP-11): production, characterization and sequencing. Minerva Biotecnologica. pp. 175-182. ISSN 1120-4826

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Aim. Kinetoplastid membrane protein-11 expresses life cycle stages of all kinetoplastidae parasites. Previous studies have demonstrated that kinetoplastidae KMP-11 gene is highly conserved and may be useful for vaccine strategies against Leislunaniasis. In this study, we isolated Leishmania major (MRHO/IR/75/ER) KMP-11 gene and formulated a pcKMP-11 recombinant expressing plasmid as a candidate DNA vaccine against cutaneous Leishmaniasis. Methods. After gene amplification, KMP-11 fragments were cloned into pTZ57R/T standard cloning vector and transformed in E. coli, then subcloned into pcDNA3 eukaryotic expression vector and pcKMP-11 recombinant plasmid was transfected to CHO eukaryotic cells. Amplification, sequencing, cloning and transfection of gene were performed successfully. mRNA transcription of KMP-11 gene in CHO cells was confirmed by RT-PCR methods. Results. Sequence results were compared with other records of kmp-11 in gene bank and a 97-99 identity was showed. Comparison of KMP-11 protein with other records showed that this protein have 92 amino acids. Additionally, a silico analysis of 3D structures of the wild type and double mutant KMP-11 proteins show that the mutations in position 16 and 41 have led to a change in structure conformation and stability. Conclusion. Present results show that KMP-11 can be an excellent candidate for immunization against leishmaniasis.

Item Type: Article
Keywords: Kinetoplastid membrane protein-11 (KMP-11) Leishmania major Leishmaniasis, cutaneous Vaccines, DNA kinetoplastid membrane protein-11 trypanosoma-cruzi molecular characterization african trypanosomes immune-response kmp11 antigen donovani protection Biotechnology & Applied Microbiology
Page Range: pp. 175-182
Journal or Publication Title: Minerva Biotecnologica
Journal Index: ISI
Volume: 26
Number: 3
ISSN: 1120-4826
Depositing User: مهندس مهدی شریفی

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