关键词: 意大利蝗, 血蓝蛋白亚基Ⅰ, 基因克隆, 表达分析, 原核表达

Abstract: Hemocyanins are copper-containing (Cu2+) proteins in vivo of insect and play important roles not only in respiratory  but also in energy storage, antibiosis and antivirus. To investigate the structure and expression profiles of Calliptanus italicus hemocyanin subunit typeⅠ(CitHc-1), the complete cDNA (2 335 bp) of CitHc-1 was obtained by RACE method. The CitHc-1 contained an open reading frame of 2 079 bp encoding 692 amino acids. The predicted molecular weight is 79.88 kDa. Sequence alignment revealed that the CitHc-1 shared an identity of 91%~94% with hemocyanins from other Acridoidea species. The dendrograms of the arthropod hemocyanin subunit typeⅠinferred by Neighbor-joining (NJ) method showed that, Calliptanus italic hemocyanin subunit typeⅠand Locusta migratoria hemocyanin subunit typeⅠformed a subclade. In order to further study the structure and function of hemocyanin subunit typeⅠfrom Calliptanus italicus, the CitHc-1 prokaryotic expression vector was established successfully and the recombined plasmid was transfected into BL21(DE3). The result of SDS-PAGE showed that the molecular weight was consistent with the theory molecular weight (32 kDa). The results provided a theoretical basis for further analysis on the function of CitHc-1 in C. italicus.

Key words: Calliptanus italicus, hemocyanin subunitⅠ, gene clone, expression analysis, prokaryotic expression