[Construction of the enhanced green fluorescent protein eukaryotic expression vector carrying CagA of Helicobacter pylori].
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.


CONCLUSIONS
Construction of the enhanced green fluorescent protein eukaryotic expression vector carrying CagA was successful.
Feng
Bai-sui
BS
Department of Digestive Medicine, Institute of Clinical Medical Research of Universities Henan China, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China. bs63@163.com
Yao
Xue-hua
XH
Zhao
Guo-qiang
GQ
D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D016480:Helicobacter pylori / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C080866:cagA protein, Helicobacter pylori; D049452:Green Fluorescent Proteins