Construction and identification of pEGFP-N1-ALDH1A1 eukaryotic expression vector
WU Chengli
WANG Yongmei
WANG Gefang
YU Guanzhen
WANG Jiejun
Abstract：Objective To construct the pEGFP?N1?ALDH1A1 eukaryotic expression vector and evaluate its biological function in human gastric cancer cell line MKN?28. Methods A cDNA encoding ALDH1A1 was cloned by gene cloning techniques;The pEGFP?N1?ALDH1A1 was constructed by use of recombinant DNA technique and was demonstrated by restriction endonuclease mapping and sequencing.The pEGFP?N1?ALDH1A1 was transfected into human gastric cell line MKN?28 by using lipofectamine 2000. The protein expression of ALDH1A1 was detected by the Western blotting assay. The activity of ALDH was detected with ultraviolet?vis?ible light detector. Results A cDNA encoding ALDH1A1 was successfully cloned;Western blotting showed the level of protein expres?sion of ALDH1A1 in gastric cancer cell line MKN?28 transfected with pEGFP?N1?ALDH1A1 were significantly increased,and the AL?DH1A1 activity in MKN?28 transfected with pEGFP?N1?ALDH1A1 were significantly increased. Conclusion The recombinant plasmid of pEGFP?N1?ALDH1A1 was constructed corrected and the protein of ALDH1A1 overexpressed in human gastric cancer cell line MKN28. It provided solid experiment foundation for the further studies on the function of ALDH1A1 in gastric cancer.
Keywords：Gastric cancerALDH1A1Eukaryotic expression vectorCell transfection
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 865-869 )