Expression and purification method of FXYD domain-containing ion transport regulator 6 (FXYD6) in intracellular region

Abstract

The invention discloses an expression and purification method of a FXYD domain-containing ion transport regulator 6 (FXYD6) in an intracellular region. The expression and purification method utilizes a primer pair shown in nucleotide sequences of SEQ ID No: 2 and SEQ ID No: 3. Through the PCR technology, a FXYD6 sequence in an intracellular domain is amplified. The amplified FXYD6 sequence in an intracellular domain is inserted into an expression vector pGEX-6P-1 so that a recombinant vector pGEX-6P-1-FXYD6 in an intracellular domain is constructed. The recombinant vector pGEX-6P-1-FXYD6 is transformed into a host cell and expresses a FXYD6 intracellular recombinant protein in the host cell. The expressed FXYD6 intracellular recombinant protein is identified, then is purified by an affinity column, and then is subjected to SDS-PAGE analysis-based purity identification. The purified FXYD6 intracellular protein obtained by the expression and purification method can be further used for preparation of a monoclonal antibody of the FXYD6 in an intracellular domain. The expression and purification method provides a beneficial tool for further research on FXYD6 tissue distribution.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the expression and purification of FXYD6 intracellular domain protein. Background technique [0002] The FXYD6 gene is located on the long arm of chromosome 11 (11q23.3). The FXYD6 protein is a new member of the FXYD family, and its full name is FXYD domain-containing ion transport regulator 6 (FXYD domain-containing ion transport regulator 6). It belongs to a single transmembrane protein, and its structure contains a single transmembrane fragment with an extracellular fixed FXYD sequence, which is an ion channel or an ion channel regulator. FXYD6 protein is distributed as a functional region of a membrane protein: the signal peptide is located at 1-18aa, the extracellular region is located at 19-38aa, the transmembrane region is located at 39-56aa, and the intracellular region is located at 57-95aa. [0003] FXYD6 has a great effect on the excitability of neurons in the...

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Problems solved by technology

[0004] There are few reports on the relationship between FXYD6 and tumors, but the mutation or deletion of FXYD family is likely to cause serious pathological changes

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