Human CD44 gene eukaryotic expression vector and expression in breast cancer cells.docVIP

 PAGE \* MERGEFORMAT 18 Human CD44 gene eukaryotic expression vector and expression in breast cancer cells Study: Housing New XU Wen-lin Xu Qian Zhang Hui Chen Chen Chiao-Yun Chen Fang Lili [Abstract] Objective: The human breast cancer doxorubicin-resistant cell lines (MCF 7/Adr) Construction of cloned CD44 gene and CD44 gene eukaryotic expression vector pcDNA3.1 CD44; will pcNDA3.1 CD44 stably transfected into human breast cancer cell line (MCF 7 cells. Methods: MCF 7/Adr cells were extracted RNA RT PCR, to obtain full-length cDNA template, which will contain CD44 gene cDNA template of double restriction enzyme digestion obtained with the CD44 gene restriction sites, restriction sites and then with CD44 gene was sequenced after TA cloning and then subcloned into the eukaryotic expression vector pcDNA3.1 in again after double restriction analysis sequencing . liposome pcDNA3.1 CD44 transfected into MCF 7 cells 48 hours after RT PCR and flow cytometry after transfection of CD44 gene and protein expression in MCF 7 cells. Results: Success from the people MCF 7/Adr cloned cells and the CD44 gene eukaryotic expression vector transfected MCF 7 was detected in the CD44 gene mRNA and protein expression, which will be the CD44 gene and protein expression of transiently transfected MCF 7 cells were After two weeks of G418 selection to obtain positive clones. Conclusion: The establishment of human CD44 gene cloning and eukaryotic expression plasmid, pcDNA3.1 CD44 can be expressed in MCF 7 cells in MCF 7/Adr a newly discovered gene variant CD44 (Gene Bank No. FJ216964), which further study the gene function for the foundation. [Keywords:] CD44, multidrug resistance, invasion, breast cancer [Abstract] Objective: To clone CD44 gene from multidrug resistant human mammary carcinoma cells (MCF 7/Adr) and construct its eukaryotic expression vector. Which was transfected into MCF 7 cells to obtain position clone.Methods: The full length cDNA encoding CD44 was obtaine