Research On The Mechanism And Function Of RNA Binding On The Surface Of Saccharomyces Cerevisiae Spore

Saccharomyces cerevisiae,as a commonly used model strain,has extensive research and application value.Saccharomyces cerevisiae cells exist as vegetative cells under nutrient-rich conditions,and their cell walls are mainly composed of glucan and mannan layers;when nutrients are lacking,diploid yeast will enter a dormant state and produce haploids Spores,the spore wall is composed of mannose layer,glucan layer,chitosan layer and dityrosine layer from the inside to the outside.Among them,the special structure of the chitosan layer and the dityrosine layer enables the spores to resist external environmental pressure.However,so far,the assembly mechanism of the spore wall of Saccharomyces cerevisiae is still unclear.In the early research in the laboratory,it was found that Saccharomyces cerevisiae spores can be swallowed by mammalian cells HEK293 T,and the phagocytosis is related to the surface RNA of the spores.Based on this discovery,this research carried out the following three explorations: RNA is found on the spore wall,knock out genes related to RNA on the surface of spores,these genes affect the surface RNA of the spore and the structure of the spore wall;extracting and purifying RNA from the surface of Saccharomyces cerevisiae spores,exploring the function of RNA on the surface of spores;using spore wall materials,establishing the delivery of micron-sized particles to non-phagocytosis Cellular method.The main research results are as follows:(1)In Saccharomyces cerevisiae,surface related genes SIP18 and GRE1 were knocked out,and the amount of RNA on the surface of the cell spores was higher than that of the wild type.At the same time,it was found that the knockout of SIP18 made the spore wall dityrosine content,RNA rebinding ability and Sub2 protein in the spore wall.The binding capacity of the spore surface is lower than that of the wild-type spore.This proves that Sip18 protein has a protective effect on the protein on the spore wall of Saccharomyces cerevisiae.(2)Saccharomyces cerevisiae spores treated with RNase have a lower survival rate than wild-type spores and RNA can protect the spores.(3)Electrostatic adsorption and chemical cross-linking were used to co-incubate the surface molecules of the spores with latex beads(diameter 2 ?m)with amino groups on the surface.The study found that the surface molecules of the spores can increase the phagocytosis efficiency of the latex beads by HEK293 T cells.This study conducted a preliminary study on the functions of surface proteins and RNA of Saccharomyces cerevisiae spores,and laid a foundation for the study of the mechanism of non-macrophage phagocytosis of large particles and the functional characteristics of surface molecules of Saccharomyces cerevisiae spores.The research results provide a theoretical basis for the future medical application of Saccharomyces cerevisiae spores and the establishment of a new drug delivery system for micron materials.