Construction of a eukaryotic expression vector of the gene encoding rat interferon-gamma-inducible protein and its expression in NIH 3T3 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 615-618, 2009.
Article
在 Zh
| WPRIM
| ID: wpr-233728
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct an expression vector of the gene encoding rat interferon-gamma-inducible protein (IP-10) and identify its expression in NIH 3T3 cells.</p><p><b>METHODS</b>IP-10 gene was amplified by polymerase chain reaction (PCR) and inserted into the eukaryotic expression vector pcDNA3.1(+). After identification by PCR, restriction endonuclease digestion and sequence analysis, the recombinant expression vector pcDNA3.1(+)-IP-10 was transfected into NIH 3T3 cells via liposome. Immunofluorescence method was used to confirm the expression of pcDNA3.1(+)-IP-10 in the transfected NIH 3T3 cells. The expression of IP-10 protein in the supernatant of the transfected cells was examined by Western blotting.</p><p><b>RESULTS</b>PCR, restriction endonuclease digestion and sequence analyses confirmed successful construction of the recombinant vector pcDNA3.1(+)-IP-10. Immunofluorescence assay identified the expression of pcDNA3.1(+)-IP-10 in NIH 3T3 cells, and the expression of IP-10 protein was detected by Western blotting in the supernatant of the transfected cells.</p><p><b>CONCLUSION</b>A eukaryotic expression vector pcDNA3.1(+)-IP-10 has been successfully constructed, which provides the basis for investigating the therapeutic effect of IP-10 on Th1 type autoimmune disease.</p>
全文:
1
索引:
WPRIM
主要主题:
Plasmids
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Transfection
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DNA Restriction Enzymes
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Gene Expression
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Fluorescent Antibody Technique
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NIH 3T3 Cells
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Chemokine CXCL10
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Genetic Vectors
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Genetics
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Metabolism
限制:
Animals
语言:
Zh
期刊:
Journal of Southern Medical University
年:
2009
类型:
Article