[Construction of the eukaryotic expression vector of human syk and its effect on MHC-I expression of human breast cancer cells]
| Autor: | Bao-en, Shan, Xiao-tian, Song, Yi-xin, Qi |
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| Rok vydání: | 2008 |
| Předmět: |
Base Sequence
Reverse Transcriptase Polymerase Chain Reaction Genetic Vectors Intracellular Signaling Peptides and Proteins Seminal Plasma Proteins Nuclear Proteins CHO Cells Protein-Tyrosine Kinases Flow Cytometry Transfection Cancer Vaccines Interferon-gamma 4-1BB Ligand Cricetulus Cricetinae Animals Humans Syk Kinase Cells Cultured Plasmids |
| Zdroj: | Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 24(1) |
| ISSN: | 1007-8738 |
| Popis: | To construct the eukaryotic expression vector of human spleen tyrosine kinase (Syk) and study the effect of Syk on MHC-I expression of human breast cancer cells.A CDS fragment of human syk was obtained by RT-PCR from human breast cancer cells MDA-MB-468 and then the fragment was cloned into the expression vector pcDNA3.1D/V5-His-TOPO. After restriction endonuclease digestion, PCR and DNA sequencing confirmation, the recombinant hsyk expression plasmid was transfected into human breast cancer cells MDA-MB-231 by using lipofectamine protocols. After G418 selection, the cells stably expressed Syk. The expression of Syk, MHC-I and ICAM-I in the transfected cells was detected by Western blot, RT-PCR and flow cytometry (FCM).The eukaryotic expression plamid pcDNA3.1D/V5-His-TOPO/hsyk was constructed and it was expressed in the human breast cancer cells MDA-MB-231. The MDA-MB-231/Syk cells highly expressed MHC-I and ICAM-I.The successful construction of eukaryotic expression plamid pcDNA3.1D/V5-His-TOPO/hsyk and its expression in eukaryotic cells will be useful for further study of tumor immunity. |
| Databáze: | OpenAIRE |
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