Clone, expression and use of Schistosoma Japonicum signal transduction protein Sjwnt-4 gene

Abstract

The invention discloses a Wnt family gene which is amplified for the first time from Schistosoma japonicum 19-day schistosomulum by utilization of RACE technology, wherein, sequence analysis indicates that: a complete coding frame of the gene comprises 1311bp, 436 amino acids coded; and theoretical molecular weight of 49.6kD. Homology analysis results indicate that: amino acid sequences of the gene have typical Wnt family protein characteristics; similarity of the amino acid sequences with amino acid sequences of Dugesia japonica and human Wnt4 is relatively 43 percent and 37 percent; the gene is presumed to be a Wnt4 gene of schistosome and named as Sjwnt4 (GenBanK Log-On No.: DQ643829). Real-time quantitative PCR analysis indicates that: the gene is expressed all in 14-day schistosomulum, 19-day schistosomulum, 31-day imago, 44-day female worms and 44-day male worms. The invention constructs a pronucleus expression vector pGEX-4T-2-Sjwnt4 of the gene and applies an Escherichia colisystem for expression; expression proteins exist in the form of inclusion bodies; Western blottings indicate that expression products can be identified by crude antigen immune serums of Schistosoma japonicum imago.

Description

Technical field: [0001] The invention belongs to the field of gene technology, and in particular relates to the cloning, expression and application of a Japanese blood sucking signal transduction protein Sjwnt-4 gene. Background technique: [0002] Wnts are a kind of secreted glycoproteins that can be found in hydra, insects and vertebrates. After being secreted by cells, they bind to membrane receptors of themselves or adjacent cells, stimulate downstream signaling pathways, and regulate the expression of target genes in the nucleus. Expression, determines the differentiation fate of cells, and plays a broad regulatory role in animal development. Abnormal spatiotemporal expression or abnormal activation of Wnt protein is often related to the occurrence of tumors. Studies have shown that the Wnt signaling pathway plays a key role in regulating the normal development of the mammalian reproductive system. It is mainly involved in the formation of Mullerian ducts and their der...

AI Technical Summary

Problems solved by technology

But so far, little research has been done on the signal transduction system of schistosome growth and development, resulting in slow progress in the development of highly effective candidate vaccine molecules and the screening of new drugs

However, there are no research reports on the Wnt gene of schistosomiasis at home and abroad.

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