Abstract: [Abstract] Objective To construct the eukaryotic expression vector of SOD1-G93A gene. Methods The M1 and including mutation gene of M2 were amplified by PCR. The two amplified gene sequence was annealed to form a chimeric SOD1-G93A gene with gene splicing by overlap extension. The resulting chimera was cloned in pcDNA3.1(-) vector and verified by sequencing analysis. Results The result showed that SOD1-G93A gene was successfully amplified with gene splicing by overlap extension, and sequencing was not changed except one base compared with the GeneBank. The recombinant plasmid SOD1-G93A-pcDNA3.1(-) was identified by double digestion and sequencing results entirely consistent with the expected. Conclusion The eukaryotic expression vector of SOD1-G93A-pcDNA3.1(-) was successfully constructed.