Cloning of Wnt Fz5 encoding gene of Schistosoma japonicum and its differential expression during different developmental stages / 中国血吸虫病防治杂志

ABSTRACT

Objective To clone full length gene sequence containing open reading frame (ORF) of Schistosoma japonicum SJCHGC08304, analyse its category encoding Wnt protein, and to understand the mRNA expression dynamics of the gene during different developmental stages. Methods Based on a EST identified in Genbank, the full length gene of SJCHGC08304 was cloned with RACE technique from 7 d hepatic schistosomula and 42 d adult worms, and its complete ORF was analysed by bioinformatics tools. The expression of the gene during different developmental stages was analysed by using Real-time fluorescent quantitative PCR technique. Results A new gene was obtained, containing a complete ORF with 2 604 nucleotides, and encoding 867 amino acid with 98.264 kD of molecular mass. Real-time fluorescent quantitative PCR showed that the mRNA level of this gene was the highest in the 18 d schistosomula, followed by 14 d schistosomula, 42 d male worms, 32 d adult worms, egg, 23 d adult worms, 27 d adult worms, respectively, and there were no expression of the gene in 42 d female worms. Conclusions The gene has a typical characteristics of Wnt frizzled family proteins, the similarity is best to Xenopus laevis. By comparison analysis to speculate as Fz5 gene, we denominate this gene as "sjFz5" (GenBank accession No. EU370926). SjFz5 has differential expressions during different developmental stages.