Bioinformatics analysis , eukaryotic expression and identification of human gastric intrinsic factor
10.19405/j.cnki.issn1000-1492.2025.08.017
- VernacularTitle:人胃内因子的生物信息学分析 、真核表达与鉴定
- Author:
Chengyi Fan
1
;
Hongyan Jiang
2
;
Bo Wang
2
Author Information
1. Hainan Afiliated Hospital of Hainan Medical University/Hainan Provincial People ′s Hospital , Haikou 570311 ; Chongqing Three Gorges Medical College , Chongqing 404120)
2. Hainan Afiliated Hospital of Hainan Medical University/Hainan Provincial People ′s Hospital , Haikou 570311
- Publication Type:Journal Article
- Keywords:
gastric intrinsic factor;
eukaryotic expression;
bioinformatics;
protein purification;
activity identifica⁃ tion;
ELISA
- From:
Acta Universitatis Medicinalis Anhui
2025;60(8):1485-1490
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze the physical and chemical properties of human gastric factor by bioinformatics method , and to express and purify the protein.
Methods:Protein online analysis software was used to predict the physical and chemical properties of gastric intrinsic factor and analyze their hydrophilicity and hydrophobicity. The online tool was used to predict and analyze the human gastric factor signal peptide and its subcellular location . The eukaryotic expression recombinant plasmid pcDNA3 . 1 ( + ) _human GIF_His tag was constructed , and the gastric intrinsic factor was purified by nickel column after expression in HEK293F cells . The purity and activity of purified gastric intrinsic factor were verified by SDS_PAGE , Western blot and indirect ELISA .
Results:Gastric factor contained 417 amino acids and was a hydrophilic acid stable protein . It was a secreted protein with Sec original signal peptide . pcDNA3 . 1 ( + ) _human GIF_His tag recombinant plasmid was successfully constructed and soluble expression was obtained in HEK293F cell expression system.
Conclusion :The eukaryotic source of human gastric intrinsic factor is successfully prepared , and the bioinformatics results show that the protein is a hydrophilic acid stable secreted protein , laying a foundation for the subsequent use of this protein as an immunogen and protein calibrator to construct an immunoassay for gastric factor.
- Full text:2026041111585340342人胃内因子的生物信息学分析、真核表达与鉴定_范成意.pdf
