The Studies On Cloning And Expression Of Cu/Zn-SOD In Branchiostoma Belcheri Tsingtauens

This article using rapid amplification of cDNA ends?RACE?,the full-length cDNA of copper-zinc superoxide dismutase gene was cloned from Branchiostoma belcheri tsingtauense in this paper.Its full-length cDNA is 1258 bp,including a 468 bp open reading frame?ORF?encoding 155 amino acids,which has seven Cu-Zn binding sites.The molecular weight deduced is 15718.32 Da and isoelectric point is5.60.There is no subcellular localization of signal peptide and transmembrane domain.The encoded protein belongs to icCu/Zn-SOD.The analysis of plylogenetic tree showed that the icCu/Zn-SOD from Branchiostoma belcheri tsingtaunese was located between vertebrate and invertebrate,had close relatives with the vertebrate.In situ hybridization results showed that the Cu/Zn-SOD are expressed in the dorsal nerve,hepatic caecum and gonad.We thought it is because these organizations constitute the important parts of the amphioxus antioxidant immune system.And Cu/Zn-SOD is considered most important kind of enzymea family of superoxide dismutase?sod?family.So this gene expressed in these parts of a large number can help amphioxus normal antioxidant immune response.Using Quantitative real-time PCR to detection the different of Cu/Zn-SOD's expression in Branchiostoma belcheri tsingtauens which was raise with different concentration of Cu²⁺.The results showed that based on 0.01mg/L,when the concentration is 0.03mg/L the expression of this gene is relatively high.Then decreased expression.When the concentration is 0.07mg/L the expression of this gene is relatively less.