Study Of Genome, Transcriptome And Wnt Gene Family Of Taenia Solium

Taenia solium is belongs to platyhelminth. The adult tapeworm fosters in the intestine of human and its larvae infect both humans and pigs, parasitizing in brain, eye and muscle etc, where causes cysticercosis. Neurocysticercosis can be developed once the larvae infected the brain of human, which causes the central nervous system symptoms, such as epilepsy, and even death. The disease is distributed worldwide in many developing countries in the Central America, Latin America, Southeast Asia and the Saharan desert surrounding regions in Africa. In China, the disease is enzootic in the rural areas in the Southwest, and it directly threatens the food safety and people healthy. Lack of the basic knowledge has limited the applied research for solving the practical issues. We designed genome project of T. solium in order to reveal the complicated life of this parasite.In the present study, the combined technology of454and Solexa were used to de novo sequence the genome of T. solium. The draft genome map was drafted, in which the genome coverage was45times, and the genome size was131Mb. In annotation,11,902genes were predicted. In addition, through sequencing of activated and non-activated cysticerci using RNAseq technology,2,766genes were found, of which1,904(68.8%) was upregulated and862(31.2%) were downregulated in the activated cysticerci. The significant regulation of the Wnt family and its signal pathway were revealed, and6members of the Wnt family were found, in which Wnt4and Wnt11b were significantly upregulated in the activated cysticercus.The full length genes of the Wnt family were obtained through RT-PCR,5’-and3’-RACE amplification. The full length Wntl was3,316bp containing two transcripts. The ORF of Wntl-1was1599bp containing6exons and the ORF of Wntl-2was1515bp containing5exons. The Wnt2was21,355bp with an ORF of1,746bp, which contains6exons. The Wnt4was10,741bp, in which the various transcripts were present at the5’terminus, but thier ORFs were exactly the same with the size of1,473bp and consisting of4exons. The Wnt5was12,272bp with an ORF of1,095bp containing5exons. The Wntl la was3,746bp with an ORF of1,263bp, which comprised of6exons; the Wntllb was14,537bp with an ORF of1,608bp, which comprised of6exons as well. Through bioinformatics analysis, it was found that all of the Wnt genes possessed signal peptides, disulfide bond, many conserved cysteine, asparagines and glycocine sites, N-glycosylation, phosphorylation and myristoylation. A phylogenetic tree was inferred using amino acid sequences of Wnts from various species, which demonstrated that the Wntl from platyhelminthes was not placed onto the same cluster with the other species. The tree also revealed the close relationship of Wnt2, Wnt4; Wntl la and Wntl lb, and the loss of wnt genes in the platyhelminthes. The loss of the wnt genes is likely because of the adaptation of the parasite in its mammal hosts.GAPDH as internal control, qPCR was applied to investigate Wnt4and Wntl lb in the activated and non-activated cysticerci, shown they were all upregulated in either classical pathway LEF1or non-classical pathway NFAT. In situ hybridization was down using Wnt cRNA probes, in which the antisense probe gave a signal at the tail of the activated cysticercus. The results indicated that the Wnt4might participate in from the larvae to the adult development through the classical signal pathway.