Study of human ZnT-8 as novel immunomodulator for type 1 diabetes mellitus
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Study of human ZnT-8 as novel immunomodulator for type 1 diabetes mellitus
Journal of Chongqing Medical UniversityVol. 40, Issue 7, Pages: 955-959(2015)
作者机构:
1. 中山市人民医院内分泌科,广东,528400
2. 第三军医大学第一附属医院内分泌科,重庆,400038
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Published:2015,
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Liu Baoying, Jiang Youzhao, Liang Ganxiong, et al. Study of human ZnT-8 as novel immunomodulator for type 1 diabetes mellitus. [J]. Journal of Chongqing Medical University 40(7):955-959(2015)
DOI:
Liu Baoying, Jiang Youzhao, Liang Ganxiong, et al. Study of human ZnT-8 as novel immunomodulator for type 1 diabetes mellitus. [J]. Journal of Chongqing Medical University 40(7):955-959(2015)DOI:
Study of human ZnT-8 as novel immunomodulator for type 1 diabetes mellitus
Objective:To lay a foundation for screening novel immunomodulator for type 1 diabetes mellitus by generating human Zinc transporter 8(of ZnT-8,a novel type 1 diabetes autoantigen) recombinant protein using a prokaryotic expression system. Methods:The COOH-terminal cDNA of the human ZnT-8 gene was subcloned into the pET-32a vector and transformed into competent Es-cherichia coli(E.coli) BL21 cells. After transformation E.coli was induced using isopropyl β-D- 1-thiogalactopyranoside;human re-combinant of ZnT-8 protein was extracted,purified by Nickel sepharose affinity chromatography,and was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. Forty female non obese diabetic(NOD) mice of four weeks old were di-vided into 2 groups in random. In ZnT-8 recombinant protein group,mice were given recombinant protein of ZnT-8;in control group,mice were given saline. The body mass,blood glucose levels and serum C-peptide levels were recorded every week. Results:The cD-NA of the carboxyl terminal of ZnT-8(306 bp) was successfully amplified by PCR. The sequence matched completely with the se-quence of ZnT-8-COOH in GeneBank. The recombinant ZnT-8-COOH was successfully expressed in a soluble form in E.coli BL21 and purified. Compared with those of control group,the body mass in ZnT-8 recombinant protein group was higher at 28 weeks old(P<0.05);the glucose levels in ZnT-8 recombinant protein group were lower at 28 weeks old(P<0.05);the serum C-peptide levels in ZnT-8-COOH group were higher at 28 weeks old(P<0.05). Conclusion:Human recombinant of ZnT-8-COOH protein is successfully prepared. The recombinant protein can be used as immunomodulator in type 1 diabetes mellitus.