Abstract:
Objective To establish the quantitatively inducible PRDM1-expressing stable cell line in order to create a platform for the study of PRDM1 function.
Methods Full-length PRDM1 cDNA was inserted into the multiple cloning site of pMEP4 vector.A pMEP4-PRDM1 expression vector was instantaneously transfected into SALT3 cell line by electroporation,followed by hygromycin B selection.Hygromycin-resistant stable clones were expanded and PRDM1 protein expression was detected at different doses of CdSO4 by Western blot.
Results The pMEP4-PRDM1 expression vector was successfully constructed and subsequently transfected into the SALT3 cell line under optimal electroporation conditions.Six hygromycin-resistant stable clones were established and expanded.The PRDM1 protein expression in these stable cell lines was proved to be quantitatively correlated with the dose of CdSO4.
Conclusion Quantitatively inducible PRDM1-expressing stable cell line can be established using the pMEP4 vector,thus laying a good foundation for further analysis of the function of this gene.