Abstract:

BACKGROUND: Overexpression of angiotensin-converting enzyme 2 (ACE2) by transgene treatment is a tendency on curing angiocardiopathy. The construction of ACE2 gene expression plasmid can provide tools for those relative researches.
OBJECTIVE: To construct eukaryotic expression vector containing mice ACE2 gene and to verify its protein expression.
METHODS: The full-length cDNA of mice ACE2 was amplified from mouse kidney by RT-PCR, and cloned into pcDNA3.1/Hygro(+) to construct eukaryotic expression vector pm-ACE2 containing mouse ACE2 gene, and identified by enzyme digestion and sequencing. The constructed pm-ACE2 was transfected into COS7 cell by Lipofectamine 2000, and its expression in eukaryotic cell of pm-ACE2 was detected by Western-Blot.
RESULTS AND CONCLUSION: Mice ACE2 gene was successfully cloned and linked to vehicle pcDNA3.1/Hygro(+). Sequencing result showed that there were three spots of samesense mutation. The pm-ACE2 expressed ACE2 protein in eucaryon. All findings demonstrated that a mouse ACE2 eukaryotic expression vector is constructed successfully, which can express ACE2 protein.