| Mitochondrion is one of the deplicature organelle with non-nuclear genome materials in eukaryotic cells.Mitochindrion are essential for cell respiration,biosynthesis of certain metabolites,ion homeostasis and apoptosis.Although most of the genes from mitochondrion transferred into the nuclear genome during the evolute process.This study would explore the evolute mechanism of mitochondrial genome by analysising the Saccharomyces sensu stricto mitochondria genome and designing the minimum Saccharomyce mitochondrial genome.The contents of this study as followed:(1)Isolation of Mitochondria from the yeast Saccharomyces mikatae and Saccharomyces kudriavzevii by differential centrifugation.Further,the mitochondrial genome could obtain though that the mitochondria destoried by 0.1%SDS reagent and the mitochondrial genome purified by.phenol-chloroform extraction.(2)The entire Saccharomyces mikatae and Saccharomyces kudriavzevii mitochondrial genome from the previous step were sequence by Illumina sequencing olatforms and assembled by SOAPdenove.The mitochondrial genome sizes of Saccharomyces mikatae and Saccharomyces kudriavzevii were 73 KB and 72KB,respectively.(3)The mitochondrial genomes of Saccharomyces mikatae and Saccharomyces were annotated based on all mitochondrial genes in Saccharomyces cerevisiae.The locatin of tRNAs,rRNAs and introns were detected by tRNAscan,RNAweasel and MFannot.(4)The phylogenetic tree was build by MEGA based on the fifteen yeasts mitochondrial genomes which were obtained from sequencing and download.Further,the functional genes,the structure of genomes,the location of introns and the mechanism of evolution would be analyzed.(5)The proterties of ancestral mitochondrial genome were obtained by genomic comparison and evolution analysis.The conservation of fuctional region and non-fuctional region would be studied.Finally,the minimum mitochodrial genome was designed and synthesized by cut down the non-fuctional region and introns. |
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