MRNA Sequencing And Transcriptome Characteristic Of Echinococcus Granμlosus

Objective: the expression profile database of protoscolex is established preliminarily bymRNA sequencing of protoscolex and analysis of expression profile to comprehend geneexpression and protein composition of protoscolex，lay the foundation for the research ofrelationship between the parasites and hosts. it woμld be the theoretical basis of finding thenew methods of diagnosis， screening new drug targets and selecting the vaccine candidateMethods1Sample preparation: TRIZOL method was used to extract total RNA fromhuman protoscolex;2Illumina’s solexa sequencing platforms for RNA sequencing:○1Fragmentation of the whole transcriptome RNA： the oligo(dT) was used toselect poly(A)RNA.then the RNA was fragmented and cleaned up.the yield and sizedistribution of fragment RNA was assessed.○2Amplified library construction: reversetranscription was performed and the cDNA was purified.then we amplified the cDNA，performed size selection and purified the amplified DNA. Finally，we assessed the yield andsize distribution of amplified DNA.○3solexa system templated bead preparation andsequencing.○4Clean reads was obtained from sequencing resμlt and assembly-first methodwas used to get unigene.3.Gene annotation and the analysis of metabolic pathways wasperformed by comparing unigene with non redundant database of NCBI， the gene ontologydatabase and KEGG database gene.Resμlts1. OD260/OD280of total RNA is1.81and the concentration is42.75μg/μl.total mass was more than200u g.it meet the requirements of the sequencing.2. We get18569contigs，the total length of the contig was71329bp and the averagelength was384bp，the smallest contig was201bp，the largest contig was4618bp. The N50was384bp. the predicted unigene was up to9029.3. Comparing the unigene with non redundant database of NCBI，the gene ontology database and KEGG database gene，we got7441annotation comparison information in NRdatabase of NCBI，10550information from gene ontology and4731annotation in KEGG.4731annotated genes from KEGG was mapped in241metabolic pathways，these metabolicpathways was associated with metabolism process，the genetic information process，environment relative processes，cellμlar processes and human disease respectivelyConclusion1.We summed up a set of total RNA extraction method suitable forprotoscolex.2.The mRNA sequencing of protoscolexes was completed and the transcriptomedatabase of protoscolexes was established preliminarily.3. Protoscolex expression profileanalysis was completed. Gene annotation and function classification was performed withbioinformatics method.4. A preliminary metabolic pathways analysis of protoscolex wasaccomplished.