Construction Of Anti-Ghrelin DNA Oral Saccharomyces Cerevisiae And Evaluation Of Its Effect

Since Steinman et al.discovered in 2001 that Saccharomyces cerevisiae can activate DCs and trigger specific cellular immune responses,researchers have begun to focus on the development of oral vaccines using recombinant yeast.Numerous studies have shown that Ghrelin plays an important role in the mediation of physiological activities such as appetite,body weight and fat metabolism,especially in promoting the body’s feeding process.Studies have shown that a decrease in Ghrelin content reduces appetite and affects the body weight of the animal,and studies have shown that the reduction of Ghrelin content in the body does not affect the growth function of the mouse.Therefore,in recent years,the idea of reducing the body weight of the animal by reducing the ghrelin content and then achieving the weight loss effect has been widely concerned.Based on the yeast immune system developed by our team,this study constructed the genomic expression vector JMB84-CMV-Ghrelin of Ghrelin gene and the eukaryotic expression vector JMB84 which expresses chicken egg albumin gene(OVA).-CMV-OVA-Ghrelin,followed by the successful construction of two oral immunized yeasts using S.cerevisiae JMY1.In the experiment,the empty vector group and the PBS group were used as the control group,and the different oral recombinant yeast groups were used as the experimental group,which were named JMB84-CMV-Ghrelin group and JMB84-CMV-OVA-Ghrelin group respectively,according to the laboratory perfect experimental scheme.After oral feeding experiments,the mice in the JMB84-CMV-OVA-Ghrelin group had the lowest daily average intake and the lowest relative growth of the body weight,indicating that the recombinant yeast can reduce the content of ghrelin in the body,and thus The mice have reduced appetite,increased energy expenditure,and reduced body weight.In addition,we have also constructed a pET32a-Ghrelin vector for prokaryotic expression.In subsequent studies,we will use the prokaryotic expression of the Ghrelin protein as an antigen to detect the antibody production in each experimental group.In summary,this study successfully constructed two eukaryotic expression vectors of Ghrelin gene,and transformed the eukaryotic expression vector into Saccharomyces cerevisiae to successfully construct two oral recombinant immune yeasts.In addition,we also performed two experiments on animal live experiments.The immune effect of recombinant yeast was studied and it was found that the immune effect of OVA-Ghrelin fusion expression was better than that of Ghrelin alone.This study provides a new way of thinking for the treatment of obesity in patients,and laid a good foundation for the further study of the system.