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Title 

Generation of expression clone set for functional proteomics of human gastric and liver cancers

Authors 

Nang-Su OhJ S ParkYeo-Jin JeonJ H OhS Y JeongJin Ok YangY W ParkHyang Sook YooNam-Soon Kim

Publisher 

Society for Experimental Biology and Medicine

Issue Date 

2009

Citation 

Experimental Biology and Medicine, vol. 234, no. 10, pp. 1220-1229

Keywords 

Functional studyGastric cancerGateway systemHigh-throughput expressionLiver cancer

Abstract 

Two thousand sixty-eight multi-purpose expression clones for the 326 candidate genes related to gastric or liver cancers were constructed using the Gateway system. These clones can be expressed as His, Glutathione-S-transferase (GST) or Enhanced version of the green fluorescent protein (EGFP) fusion proteins in E. coli, insect cells or mammalian cells. For the 246 E. coli expression clones, the GST fusion proteins had greater expression efficiency and solubility than the His fusion proteins. Approximately 20% of the expressed proteins had unexpected molecular weights. A detailed sequence analysis of these clones revealed frameshift mutations resulting from insertion, deletion or substitution of nucleotides. The results indicate that these changes in the candidate genes may affect the occurrence of gastric or liver cancers. In addition, when 105 proteins, which were expressed in E. coli at very low or undetectable levels, were expressed in insect cells, 76% of the proteins were expressed very well and most were soluble. We also found that most of the 30 proteins prepared using EGFP mammalian expression clones were localized to cellular compartments expected by Gene ontology (GO) and this localization was unaffected if the EGFP-fusion was at the N-terminal or C-terminal region of the protein. Antibody production and subcellular localization analysis of the candidate genes as well as a screen of genes involved in carcinogenesis pathways are currently in progress using these expression clones. These studies provide a valuable resource for developing a better understanding of the molecular mechanism of carcinogenesis in both gastric and liver cancer and would be very helpful in diagnosis and therapeutic predictions.

ISSN 

1535-3702

Link 

http://dx.doi.org/10.3181/0812-RM-371

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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