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Updated: Jul 15, 2026
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Published on: January 6, 2015
[Construction of the enhanced green fluorescent protein eukaryotic expression vector carrying CagA of Helicobacter
Bai-sui Feng1, Xue-hua Yao, Guo-qiang Zhao
1Department of Digestive Medicine, Institute of Clinical Medical Research of Universities Henan China, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China. bs63@163.com
Objective:
To construct a eukaryotic green fluorescent protein expressing vector containing the fragment of cytotoxin associated gene A (CagA) of Helicobacter pylori (Hp) so as to lay a foundation for the research of gene vaccine of gastric cancer. pEGFP-C3-CagA.
Methods:
The fragment of gene CagA was amplified from Hp using PCR. The amplified product was examined by electrophoresis and sequence determination. This fragment was inserted into pGEM-T plasmid and pEGFP-C3 fluorescent expression vector. The recombined plasmid pEGFP-C3-CagA was transfected into the gastric carcinoma cells of the strain BGC823 by lipoplasty method. Fluorescence microscopy was used to observe the expression of pEGFP-C3-CagA under.
Results:
CagA was inserted in the plasmid correctly. It was verified by DNA sequencing and restriction enzyme. The enhanced green fluorescent protein eukaryotic expression vector carrying CagA of Helicobacter pylori (pEGFP-C3-CagA) was recombined correctly and transfected in gastric carcinoma cell strain BGC823. Green fluorescence was observed in transfected gastric carcinoma cell.
Conclusion:
Construction of the enhanced green fluorescent protein eukaryotic expression vector carrying CagA was successful.
