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Cloning and expression of canine MMP11 protein in eukaryotic expression system Nandhakumar P.1, Mishra S.R.1, Rajoriya S.1, Karthik K.V.1, Kataria Meena1,* 1Division of Animal Biochemistry, ICAR-Indian Veterinary Research Institute (IVRI), Izatnagar, Bareilly-243122, Uttar Pradesh, India *Corresponding author: mkataria97@rediffmail.com
Abstract Mammary tumors are the leading cause for the death of female dogs in India. The major reason for the death of animal during tumor is the spread of cancer cells to distant organs. The matrix metalloproteinases (MMP) are the proteases secreted by cancer cells which cleave the extracellular matrix proteins to promote cancer metastasis. MMP 11 belongs to MMP family which is frequently reported in both human and canine cancer. The cDNA prepared using canine placental total RNA was used to amplify 1323bp MMP11 gene, which was subsequently cloned in pVAXl vector and followed by transfection into Vero cell line by Lipofectamine. The recombinant MMP11 precursor protein showed a molecular weight of 49 kDa in Vero cells confirmed by western blot and Immunofluorescence assay. The result suggests that the pVAX1. MMP11 construct may be used to screen the immunization efficiency in animal models. Top Keywords Canine, Cloning, Mammary tumors, Matrix metalloproteinases, MMP11. Top | | | |
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