Gene clone and prokaryotic expression and purification method of saddletail grouper CCR12

Abstract

Three kinds of XCR1 chemokine receptors are found in fish at present, namely XCR1, XCR1L and the CCR12, wherein the XCR1L and the CCR12 are only found in fish genomes, studies about the functions of the XCR1L and the CCR12 are few, and there is no report about fish CCR12 in-vitro expression so far. To further disclose the migration or proliferation conditions of grouper CCR12 positive cells in tissue infected by parasites, the study builds a recombinant expression vector of an N-end region and three extracellular regions of the saddletail grouper CCR12 for the first time, recombinant protein of the CCR12 is expressed through a prokaryotic system, and a foundation is laid for further in-vitro study of the grouper CCR12.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to the gene cloning, prokaryotic expression and purification of CCR12 of a grouper. Background technique [0002] Chemokines are a class of cytokines with very small molecular weight, and they have similar monomer structures (Baggiolini, 1998; Jin et al., 2005). According to their structure and the sequence of four conserved cysteines, chemokines can be divided into five types: CXC type, CC type, XC type, CX3C type and CX type (Murphy, et al., 2000; Nomiyama et al., 2008). To function, chemokines need to bind to their corresponding receptors. XCL1 is an XC-type chemokine that is produced by thymic myeloid epithelial cells, or activated T cells, NK cells, and NKT cells during homeostasis, pathogenic infection, or inflammatory response (Lei et al., 2012). XCR1 is the sole receptor for XCL1, mediated by CD8α + and CD11b - CD103 + Secreted by DC cells (Alexandre...

AI Technical Summary

Problems solved by technology

Among them, XCR1L and CCR12 are only found in the fish genome, but there are few studies on their functions, and the lack of anti-XCR1L or CCR12 antibodies also limits the isolation and function of their positive cells.

So far, there has not been any report on the in vitro expression of fish CCR12

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