Prokaryotic Expression and Antibody Preparation of Human Spermine/Spermidine N¹acetyltransferase

ObjectiveTo obtain prokaryotic expression of human spermidine/spermine N¹-acetyltransferase（SSAT）and prepare SSAT antibody. MethodsHuman SSAT cDNA was amplified by RT-PCR from total RNA of human lung cancer line A549.The SSAT cDNA was then cloned into prokaryotic expression plasmid pET15b and transformed into BL21（DE3).The expression of SSAT in host cells was induced by IPTG and recombinant SSAT protein was purified by polyacrylamide gel electrophoresis and Ni-NTA resin.The antibody acquired from rabbits immunized by purified SSAT protein were detected by Western blot and ELISA. ResultsHuman SSAT was successfully expressed in E.coli induced by IPTG and antibody of SSAT was prepared from purified recombinant SSAT. ConclusionA method for prokaryotic expression and antibody preparation of the human SSAT was successfully established.