A Study On Prolonging The Life Expectancy Of Saccharomyces Cerevisiae

For a long time,studying the mechanism of aging has been a hot spot in the field of life science research.Since the aging process is affected by various factors,such as metabolism,damage repair,and energy limitation,the mechanism of organism aging is very complicated.Higher organisms are faced with many problems such as the cost of studying aging models.Saccharomyces cerevisiae is a unicellular eukaryotic model organism.It has the following advantages as a model organism for the study of higher eukaryotes:1.Single-celled eukaryotes,2.Rapid growth and reproduction,and metabolism with short cycle,3.The genetic background is simple that genome has been completely sequenced with abundant experimental methods for genetic manipulation.Yeast genomes contain genes that inhibit the repair of yeast cell damage and regulate transcription and expression.Relieving the inhibitory effects of these genes on yeast cell damage repair can extend the life span of yeast,provide basis for studying the aging and life span of higher organisms,and build longevity and metabolism.Industrial production strains can reduce production costs and increase corporate profits.In this study,CRISPR technology was used to figure the POS5,LOS1,TEP1,GRC5,ISW2,SWI1,and YPT1 genes of S.cerevisiae.Here are some research findings:1.CRISPR/Cas9 system was used to insert the target sequence on the S.cerevisiae genome,and different types of experimental programs were designed successively.Finally,a technical route of gene insertion in S.cerevisiae genome based on CRISPR/Cas9 system was established.2.CRISPR/Cas9 system was used to edit LOS1,POS5,TEP1,GRC5,ISW2 and SWI1genes on S.cerevisiae M_2-3-3 strain genome,obtain the mutant strain named M_2-3-V.3.Site-directed mutation was conducted on the YPT1 gene of mutant S.cerevisiae strain M_2-3-V,and mutation sequence,homology arm sequence and selective marker gene sequence were spliced by overlaying and extending PCR;electric shock was carried out on the segment to transform competent cells of S.cerevisiae strain M_2-3-V,so as to screen out mutant strain and name it M_2-3-VI.4.The growth curve,replication lifespan,and chronological aging of S.cerevisiae M_2-3-3 and S.cerevisiae M_2-3-VI were determined respectively.The results showed that S.cerevisiae M_2-3-VI grew faster than original strain,the chronological aging and replication lifespan were1.5 times and 1.7 times of original strain respectively.