ZnT8 recombinant protein and preparing method and application thereof

Abstract

The invention provides ZnT8 recombinant protein and a preparing method and application thereof. The protein has the amino acid sequence shown in SEQ ID NO.1 and the nucleotide sequence shown in SEQ IDNO.2. By analyzing and screening a target segment, optimizing and encoding the sequence and a carrier, transfecting an Expi 293 eukaryocyte and stably expressing protein with antigenic activity, thepreparing method which can keep the bioactivity of natural protein and reduce the production cost is provided, and the method has wide application prospects and huge market value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a ZnT8 recombinant protein and its preparation method and application. Background technique [0002] Zinc transporter 8 (ZnT8) is a member of the SLC30 family (ZnTs protein), and the SLC30 family is responsible for transporting intracellular zinc ions to the extracellular matrix or intracellular vesicles. ZnT8 is specifically expressed in pancreatic tissue, and is mainly located in islet β cells. ZnT8 overexpression can further increase the insulin secretion of β cells in response to glucose stimulation by increasing the density of insulin secretion channels, thereby regulating the synthesis, storage and secretion of insulin. process. ZnT8 autoantibody may be an important marker of type Ⅰ diabetes, which has become an important supplement to the current diagnostic reagents. The specificity of ZnT8 in the detection of TIDM is as high as 98%, and there is still a positive rate of 37.5% ...

AI Technical Summary

Problems solved by technology

In 2004, scholars cloned the SLC30A8 gene from human islet cells for the first time, and expressed ZnT8 protein in HeLa cells. However, due to the complex molecular structure of ZnT8, it is difficult to express it in a large amount in the prokaryotic system. Now it is prepared by the cell-free expression system of reticulocytes in vitro. The main method of antigenic protein, but the content of ZnT8 protein prepared by this method is low and the cost is high, which restricts the large-scale clinical application

The main advantage of the mammalian cell and insect cell expression system is that the post-translational processing mechanism of the protein is closest to the natural form in the body, and it is easiest to retain biological activity. The disadvantage is that the expression level is usually low and the production cost is high.

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