Abstract:
In Drosophila melanogaster, the cohesion protein ORD( orientation disruptor) is required for sister chromatid cohesion, homologous recombination and synapsis in oocytes during meiosis. This study aimed to obtain the prokaryotic expression of ord from Drosophila melanogaster, and analyzed its functions by bioinformatics.The ord fragment was amplified by PCR, and the pET-sumo-ORD recombinant plasmid was constructed by homologous recombination. The His-ORD fusion protein was then induced by IPTG and identified by SDS-PAGE and Western blotting. The results show that the size of His-ORD fusion protein is about 69 kD, mainly in the form of inclusion body. Phylogenetic tree analysis suggests that the gene is closely related to the ord in Drosophila mauritiana, Drosophila sechellia and Drosophila simulans. Bioinformatics analysis shows that the predicted isoelectric point(pI) of ORD protein is 5.69, the average hydrophobicity(GRAVY) is-0.309, which is a hydrophilic protein,and its secondary structure is mainly alpha-helix and random coil. The protein interaction network analysis indicates that ORD interactes with INCENP, and colocalizes with CONA and CG13541. The results of this study will provide a theoretical and technical basis for further research on functions of this gene and facilitate the study of meiotic cohesion in Drosophila melanogaster.