Abstract:

Abstract:Objective
To construct the  CCNL1 eukaryotic expression vector pcfDNA3.1（+）-CCNL1,pVAXⅠ-CCNL1 and pEGFP-C1-CCNL1 and identify the  expressions of  CCNL1 genes in MDA-MB-231 cells.Methods The eukaryotic expression plasmid pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 were transfected into the MDA-MB-231　cells by Fugene HD Transfection Reagent.After 48 h  transfection,the differential expressions of pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 in  MDA-MB-231　cells were detected by real time PCR and Western blotting.Results After 48 h transfection,there was green fluorescence only in  pEGFP-C1-CCNL1 by  fluorescent microscope,not in pcDNA3.1（+）-CCNL1 and  pVAX1-CCNL1; the expressions of mRNA and protein from high to low were pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 recombinant vector in MDA-MB-231 cells.Conclusion The recombinant vectors pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-CCNL1  are constructed succesfully.The expression of pcDNA3.1（+）-CCNL1  in  MDA-MB-231 cells is higher.

Key words: breast , neoplasms;eukaryotic expression vector;Cyclin L1;MDA-MB-231 cells