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单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,*,*,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,微孢子虫基因组分析,服从真理，就能征服一切事物,微孢子虫基因组分析微孢子虫基因组分析服从真理，就能征服一切事物Preliminary analysis of genome of nosema bombycis潘国庆 周泽扬 向仲怀It is just the beginning of the big job,-The key lab of Sericulture of Agricultural Ministry,Southwest Agricultural University,Chongqing,P.R.China,400716.3/28/20212microsporidiaFirst recognized in 1857,till now,has been 147 years.The first genome was published 2019,which is Encephalitozoon cuniculi,with 2.9 Mb.,Found in from protists to inverbrates and all class of vertebrate.,More than 140 genera and almost 1200 species recognized.3/28/20213,But we are more than it,More researcher invest their energy into the microsporidia,especially the genus,Encephalitozoon,which is related to human diseases.,Our purpose is divided into two parts,one is for basic research,to discover the relationship between the Nb and silkworm,Nb and Ec;the other is that we are eager to find some key genes useful to prevent the Nb disease.,10/4/2024,6,background,Nosema bombycis is the first important pathogen in silkworm feeding.,Nosema bombycis test is time&energy-consuming in silkworm feeding.,10/4/2024,7,Nosema bombycis,Lineage,(full),cellular organisms,;,Eukaryota,;,Fungi/Metazoa group,;,Fungi,;,Microsporidia,;,Apansporoblastina,;,Nosematidae,;,Nosema,10/4/2024,8,10/4/2024,9,10/4/2024,10,Little information avaliable,10/4/2024,11,Proteinsentriesin NCBI,10/4/2024,12,Nb purification with Percoll,10/4/2024,13,N.b全基因组,shotgun,测序文库构建,N.b complete genome DNA,hydroshear,Kit extraction,1.5-2kb,2-2.5kb,PUC18,vector,PCR,Reaction,Forward sequence,Reverse sequence,10/4/2024,14,1.,Bascalling (Phred and Phd2fasta),2.Vector Mark(crossmatch),3.Assemble (Phrap),4.Finishing (consed),5.Repeat Mark,ORF Prediction,Gene Annotation,(,Glimmer,Blastx,Blastn,Clastalw,tRNAscan),6.GenBank(nr),测序和数据处理,10/4/2024,15,Data source,0.26 million reactions from 2.5kb genome library,the length of per reads:more than 400bp.,4000 reactions from 8kb genome library.,The total length of whole reads:6.0 X equivalent of genome,Contigs num:3000(1kb),with N50=8.0kb,Scaffold num:N50=10kb,10/4/2024,16,Preliminary result,predicted gene num:around 6,000.,The hits numbers with nr database(1e-6)is around 3,700.,The anno num:3,770,The predicted genes including intron is about 200.And the intron is small size.,The new version of the anno genes is about 5,000.,10/4/2024,17,comment,They estimated that gene num is similar with yeast.And the 2nd-version annotation was just over yesterday.And I have not get the updated data.We still need to construct a BAC library of Nb to obtain bigger scaffold,we hope we can get help from institute and university at domestic/abroad.,10/4/2024,18,Comparison Nb with other species,10/4/2024,19,Analysis of anno,3 anno from,Nosema bombycis.,(p30.4,elongation factor I,DNA repair protein);one anno from,Nosema locustae,.,2148 anno from E.c，57.0%.,97 anno from,Plasmodium falciparum,(疟原虫)3D7;63 anno from other plasmodium strains.,9 anno from,Bombyx mori,one is eIF2 ALPHA,the other are related to transposon,.,56 anno from,Salmonella enterica subsp.enterica serovar Typhi,(沙门氏菌属),;,49 anno,Salmonella typhimurium LT2;,36 anno from E.coli,52 anno from oryza sativa,171 anno form,Anopheles gambiae str.PEST,疟蚊,10/4/2024,20,Some key genes,tRNA synthetase,(,合成酶,),:,20,Two,tRNA synthetase,not found,:,asp and,tyr,Transposon,(,转位子,)and,retrotransposon,related gene:,497,Ribosomal protein:,93,Kinase,(,激酶,):,33,unknown gene:,77,ZINC FINGER:34,ENDOCHITINASE:3 items,obviously transcription related:39,obviously translation related:18,obviously,ubiquitin,related:28,obviously related to,mitochondrial,:9,10/4/2024,21,Mitochondrial related genes,10/4/2024,22,糖 酵 解 代 谢 途 径,Trehalose,海藻糖,Glucose,Trehalase海藻糖酶,Hexokinase,Glucose-6P,fructose-6P,果糖,GLUCOSE 6-PHOSPHATE,ISOMERASE,FDP,6-PHOSPHO-FRUCTOKINASE,磷酸果糖激酶,GLYCERALDEHYDE,-3-PHOSPHATE,Aldolase,醛缩酶,GLYCERALDEHYDE-3-,PHOSPHATE,DEHYDROGENASE,1.3-二磷酸甘油酸,3-磷酸甘油酸,Phospoglycerl,Kinase,?,磷酸二羟丙酮,TRIOSE PHOSPHATE,ISOMERASE,2-磷酸甘油酸,PhosphoglyceroMutase?,磷酸甘油变位酶,Enolase,烯醇酶,磷酸烯醇式丙酮酸,PYRUVATE KINASE,丙酮酸激酶,烯醇式丙酮酸,丙酮酸,10/4/2024,23,三羧酸循环代谢相关酶(E.c中未发现),succinyl-CoA synthetase beta chain（琥珀酸,-CoA,合成酶）,malate:quinone（苯醌）oxidoreductase(氧化还原酶),2-isopropylmalate synthase,3-isopropylmalate isomerase(dehydratase脱水酶)subunit,2-oxoglutarate dehydrogenase(脱氢酶)E1 component,3-HYDROXY-3-METHYLGLUTARYL CoA REDUCTASE,3-HYDROXY-3-METHYLGLUTARYL-CoA SYNTHASE 2,10/4/2024,24,PPP途径,ADP-L-glycero(甘油)-D-mannoheptose(甘露庚糖)-6-epimerase(差向异构酶),D-RIBULOSE 5 PHOSPHATE 3 EPIMERASE（D-核酮糖磷酸差向异构酶）,RIBOSE 5-PHOSPHATE ISOMERASE,5-核酮糖磷酸核糖异构酶,10/4/2024,25,Enzyme in lipid metabolism脂类代谢,Lipase(脂肪酶）,3-ketoacyl-CoA thiolase(3-酮酰基辅酶A硫解酶）,STEROL O-ACYLTRANSFERASE（固醇 O-酰基转移酶）,ACYLGLYCEROL-3-PHOSPHATE ACYLTRANSFERASE（3-磷酸甘油乙酰基转移酶）,1-ACYL-SN-GLYCEROL-3-PHOSPHATE ACYLTRANSFERASE,ACYLCOENZYME A THIOESTERASE II（脂酰辅酶硫酯酶II),10/4/2024,26,Nucleotide metabol
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