Cloning And Identification Of Echinococcus Multilocularis OCT4Gene

Alveolar echinococcosis, which is caused by Echinococcus multilocular larvae, is one of the world’s most dangerous zoonotic parasitic diseases. This is mainly caused by germinal layer cells which have strong capacities of self-renewal and differentiation, and the pluripotency of germinal layer cells play a key role in alveolar hydatid growth and life cycle.Pluripotency maintaining of germinal layer cells needs a complex regulatory system, and gene OCT4is an indispensable endogenous factor. So the further study of alveolar hydatid endogenous gene OCT4can help us explore the growth mechanism of alveolar hydatid.In this research, we got the full sequence of alveococcus OCT4homologous gene by RACE technology. Furthermore, we analysed the gene structure using bioinformatics techniques and the result showed that it contains POU domain which is of high conservation. On this basis, we conducted a series of functional assessments to protein (temporarily named EmOCT4) encoded by Echinococcus multilocular OCT4homologous gene.We obtained recombinant antibody of EmOCT4and the results of its immunohistochemistry experiments showed that EmOCT4expressed in nucleus. We introduced EmOCT4and mouse SOX2, KLF4, cMYC genes into mice fibroblasts through retro viral vector in order to reprogram cells. Finally, we successfully abtained induced pluripotent stem cells (iPSC). The result indicated that EmOC4played a significant role in mouse fibroblasts reprogramming and its function was similar to that of mouse OCT4. The experiment results provided a great help to further investigation in the role of EmOCT4in maintaining pluripotency of alveococcus germinal layer cells.