Study On Genome Sequencing And Analysis Of Taeniidae And HIV Protease Inhibitors In The Treatment Of Helminth Infections

Taenia hydatigena and Taenia pisiformis are two parasites that seriously endanger the development of animal husbandry and cause huge economic losses.Due to the lack of basic research,the mechanisms of their species adaptive evolution,host interaction,and immune evasion are still unclear,which brings great difficulties to the prevention and treatment of the disease.In this study,the genome sequencing of two tapeworms was performed for the first time,and the drafting and annotation of the whole genome was completed,and the genome characteristics of the two tapeworms were analyzed.Through the identification of drug target molecules,it was found that the tapeworm DNA-induced damage 1(Ddil-like)protein can be used as a potential drug target.The tapeworm Ddil-like protein and human immunodeficiency virus(HIV)are both retro-type aspartic proteases,with highly similar tertiary structures and active regions.In this study,the E.multilocularis EmuDdi1 protein was used to verify whether the protein is a drug target of HIV protease inhibitors.At the same time,Echinococcus multilocularis and its infected mice were used as research models to explore the possibility of HIV protease inhibitors as potential drugs for the treatment of HIV and helminth co-infection.The following results are now obtained:1.Completed the whole genome sequencing,splicing and evaluation at the chromosome level of Taenia hydatigena and Taenia pisiformis for the first time.The genome sizes are 309 Mb(Megabase,Mb)and 204 Mb,respectively;Scaffold N50 is 1.8 Mb and 20 Mb,respectively,with 11,358 and 12,097 coding genes,respectively.At the same time,complete the annotation of genome repeat sequences,coding genes and non-coding genes,as well as annotation of gene functions and signaling pathways.2.The widespread presence of endogenous virus-like sequences in the Taenia hydatigena genome was identified and analyzed.It is found that Taenia hydatigena achieves genetic diversity through alternative splicing and gene loss,and has evolved a new mechanism of immune evasion that is distinct from that of flukes.At the same time,potential drug targets such as G protein-coupled receptors,proteases,protein kinases and ion channels of Taenia hydatigena were identified.3.Phylogenetic analysis of the genome confirms the relatedness of Taenia hydatigena and Taenia pisiformis and other flat animals.Based on genome and transcriptome sequencing data,genes under positive selection pressure on the Taenia pisiformis tapeworm lineage were analyzed,and the adaptive evolutionary characteristics of the parasite to adapt to the host were discovered.4.The E.coli prokaryotic expression recombinant plasmid rEmuDdi1 was constructed,expressed and purified to obtain the recombinant protein with a size of about 45 kD.The enzyme kinetics experiments proved that the protein has hydrolytic activity under neutral pH and alkaline valence.5.Using RNA interference technology,transcriptome sequencing and in vitro drug inhibition experiments of HIV protease inhibitors on E.multilocularis and enzyme activity inhibition experiments of rEmuDdi1 protein,it is proved that the killing effect of HIV protease inhibitors on the PSCs is through inhibition mediated by the activity of EmuDdi 1.6.Using echinococcosis to infect an immunodeficient mouse model,it is proved that nelfinavir can be used as a candidate drug for HIV co-infection with helminths.