TY - JOUR ID - 58677 TI - Cloning and expression of Eimeria necatrix microneme5 gene in Escherichia coli JO - Iranian Journal of Veterinary Medicine JA - IJVM LA - en SN - 2251-8894 AU - Mayahi, Mansour AU - Jolodar, Abbas - AU - Masaeli, Sharouz - AU - Hamidinejat, Hosein AU - Seyfi Abad Shapouri, Masoud - AU - Moori Bakhtiari, Naghme - AD - Faculty of Veterinary Medicine of Ahvaz AD - Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, Iran AD - He is working at Veterinary Department of Qum State AD - Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, Iran AD - 3Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, Iran Y1 - 2016 PY - 2016 VL - 10 IS - 3 SP - 157 EP - 163 KW - Eimeria necatrix KW - gene expression KW - immunogenicity KW - microneme DO - 10.22059/ijvm.2016.58677 N2 - Background: Coccidiosis caused by Eimeria necatrix has the most economic impact onpoultry production. Micronemal proteins in Eimeria necatrix are thoughtto be critical ligands determining host cell specificity at the time ofinvasion.               OBJECTIVES: Isolation and purification of Eimeria necatrix oocysts from  Khuzestan province of Iran was performed. AcDNA encoding microneme 5 (EnMIC5) protein was cloned and expressed asrecombinant protein before the evaluation of its immunogenicity by Westernblotting. METHODS: A primer pair was designed based on the publishednucleotide sequence of Eimeria necatrix LZ strain micronem5 gene. APartial cDNA sequence fragment of 758 bp coding for microneme 5 protein(EnMIC5) was amplified by semi- Nested RT-PCR. PCR products were cloned andexpressed in a Maltose Binding protein (MBP) containing expression vector(pMAL-c2x) in Escherichia coli. The cDNA which is encoded for 252 aminoacids shows high degree of conservation. It contains the adhesive plasmapre-kallikrein and seven hydrophilic motifs. RESULTS: The results of SDS-PAGErevealed that the recombinant protein with a molecular weight of 70 kDa wasover-expressed after induction with IPTG. Western blotting results revealedthat the expressed recombinant protein was reacted with sera of the chicksinfected with Eimeria necatrix. It was suggested that this proteinshould have a good immunogenicity and can be used for further studies. CONCLUSIONS: In conclusion, the high degree of sequence homology indicates that thisprotein is immunogenic and  might be aninteresting vaccine target, and deserves further investigation UR - https://ijvm.ut.ac.ir/article_58677.html L1 - https://ijvm.ut.ac.ir/article_58677_9d39b619fb234eed44dc01914394dbc3.pdf ER -