Expression of Human CC10 Gene in Lung Adenocarcinoma A549 Cell

Objective To construct and identity human the expression plasmid pcDNA3.1-hCC10. Methods A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by RT-PCR,then inserted it into expression vector pcDNA3.1 by DNA recombinant techniques . We transfected pcDNA3.1- hCC10 in A549 cells by liposome-mediated gene transfer method. The protein expression of CC10 was detected by the techniques of immunofluorescence and Western blot. Results After the identification of restriction enzymes analysis and sequencing,the recombinant plasmidpcDNA3.1-hCC10 was confirmed which contained the correct and entire nucleotide sequence of the CC10 DNA in pcDNA3.1. The expression of CC10 in protein level ascend in A549 cells transfected with reconstructive plasmid. Conclusion The pcDNA3.1-hCC10 expression plasmid was successfully constructed, acquired stably protein expression in A549 lung cancer cell line.