Prokaryotic Expression of LINE-1 Encoded Protein L1-ORF1 and Preparation of Anti-L1-ORF1 Antibody

Aim:to detect L1-ORF1,an ubiquitous protein in tumor tissue,its polyclonal antibody with rabbits were prepared.Methods:L1-ORF1 gene was prepared by PCR amplification and cloned into pGEX-6P-1 with GST fusion.After expressing in E.coli,GST-L1-ORF1 fusion protein was purified by Glutathione Sepharose 4B and subjected to immunity with rabbits.The titer of anti-serum was measured by ELISA and the specificity was examined by Western blot with the cell lysates derived from HepG2 tumor cells.Result:The anti-L1-ORF1p antibody exhibits a high sensitivity and specificity,which was evaluated by L1-ORF1p transfected normal cells and tumor cells.Conclusion:Anti L1-ORF1p polyclonal antibody with high sensitivity and specificity were successfully acquired.