Molecular Cloning of Leishmania major gp63 Gene in BALB/c Mouse CT26 Cell Line

Abstract

Background: Leishmaniasis is now accounted as a worldwide disease, which is caused by different species of Leishmania parasite. This parasitic disease is now endemic in many areas including 16 developed and 72 developing countries. An estimated 12 million cases of leishmaniasis exist worldwide and a further 367 million are at the risk of acquiring the disease. A high rate of Leishmania and HIV coinfection has now been reported from 30 countries around the world. Th1 immune responses have the main role in eliciting immunity to Leishmania parasite. Many attempts have been made and different strategies have been approached to develop a potent vaccine against this parasite. Objectives: The main objective of this study was to clone and detect the gene encoding Leishmania major gp63 in mouse CT26 cell line. Materials and Methods:In this observational study, the Leishmania major gp63 gene segment was amplified using specific primers and then sub-cloned into pcDNA3 expression vector. The new construct was transfected into mouse CT26 cell line. Results:The results clearly showed that pcDNA3 L. major gp63 was successfully constructed and transfected into CT26 mouse cell line and these cells have a high potency in accepting and expressing Leishmania genes. Conclusions: A full length of L. major gp63 gene was cloned in to mouse CT26 cell line, which can be used in future vaccine studies for Leishmania.

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“…Then, the extracted molecules were ligated using a ligation enzyme (Promega, Durham, USA). The pcDNA3 L. major Gp63 vector was multiplied and extracted by E. coli transformation (25).…”