Bioinformatics Study On Differentially Expressed Genes Of Recombinant Yeast By Ion Beam

With the development of ion beam biotechnology,the transformation of exogenous genetic material mediated by ion implantation has become one of the technologies and methods for directed genetic breeding of plants and microorganisms.In order to further understand the influence of low-energy ion implantation mediated transgenic technology on yeast genome,transcriptome and metabolites,the research was based on the analysis of ion beam recombinant yeast which can produce quinones.In this paper,genomics,transcriptomics and metabonomic were used to study the function of differentially expressed genes of ion beam recombinant yeast N6076 and its original strain Kh08 at three fermentation time points(0 h,48 h,96 h).The results of genomics showed that compared with the original strain,the genome of ion beam recombinant yeast increased by 1993775 bp,the number of contig increased by 2,the number of genes increased by 270,the repetitive sequence,noncoding RNA of genome also changed.The results of go functional enrichment showed that in the biological process,proteins are mainly enriched in the biological regulation and metabolism process,and which can play their functions through the combination with ATP and GTP;in the cellular component,proteins are mainly enriched in cells,membranes and cell components;in the molecular function,proteins are mainly enriched in protein binding and binding activity.The results of KEGG metabolism analysis showed that compared with Kh08,the recombinant strain N6076 reduced one metabolic pathway which is ECM receptor interaction,involving one differentially expressed gene.There are metabolic pathways of ubiquinone and other terpenoid quinones,terpenoid skeleton,sesquiterpenoid and triterpenoid biosynthesis in both strains.The results were consistent with the follow-up analysis of metabolome.The results of transcriptome differential expression showed that the number of up-regulated genes was 1168 and 1069 when the recombinant strain N6076 was fermented to 48 h and 96 h,while the number of down-regulated genes was 1681 and1377 respectively;compared with 48 h,the number of up-regulated genes and down-regulated genes were 499 and 291 respectively when the recombinant strain N6076 was fermented to 96 h,while the number of original strain Kh08 was 523 and588 respectively.The results of GO enrichment showed that 18 GO functions were added to the recombinant yeast,including 14 biological processes,2 cellular components and 2 molecular functions,involving 1140 differential expression genes(DEGs).The results of KEGG metabolism analysis indicated that there were 13 new metabolic pathways in ion beam recombinant yeast,involving 77 DEGs.Among them,5 DEGs are related to the new increased terpenoid skeleton biosynthesis pathway.Mevalonate kinase(ATP: mevalonate 5-phosphotransferase,EC2.7.1.36)is the first enzyme of three consecutive ATP dependent enzymes in mevalonate pathway.It is one of the rate limiting enzymes that control the whole metabolic pathway,and also an important regulatory site in the synthesis of secondary metabolites such as terpenes and quinones.The metabonomic analysis of the two strains showed that the trend of MK gene expression in the whole fermentation process was consistent with the trend of MVA metabolic yield measured by metabonomic.In conclusion,The results of this study provide basic bioinformatics evidence for the biosynthesis of quinones and other secondary metabolites by ion beam recombinant yeast,and are verified by the results of some metabolism group,providing scientific basis for people to further understand and understand the differential gene expression of yeast mediated by low energy ion implantation and its impact on metabolism regulation.