Cdna Cloning And Prokaryotic Expression Of Ubiquitin In Fenneropenaeus Chinensis

The ubiquitin-proteasome pathway is the inportant portein degradation pathway in eucaryotic cells, and it influences many physiological activities, including immunologic system. In this paper, a partial ubiquitin cDNA sequence of Penaeus chinesis Fenneropenaeus chinensis was cloned.This cDNA sequence was 361 bp in length, containing a 234 bp open reading frame, which encoded 77 amino acid residues.The deduced amino acid sequence showed more than 95% identity with other known species ubiquitin. The deduced peptide was a predicted molecular weight of 8.69 kDa and its isoelectric point was 6.56.The most content of amino acid was leucine(11.69%),and the following ones were glutamine, isoleucine, lysine and threonine (9.09%),whitout cysteine and tryptophan.There were 11 acidic amino acid and 11 basic amino acid in it. The 3D model of ubiquitin was spheroidal,including a a-helix and twoβ-sheets,and C-terminal was a random coil,which was similar to known ubiquitin structures. Hydrophobicity analysis found the ubiquitin was hydrophillic.In this paper, the Penaeus chinesis Fenneropenaeus chinensis ubiquitin gene was cloned into prokaryotic expression vector pBV221.The recombinant plasmids were successfully introduced into E. coli BL21,and showed the ubiquitin was successfully expressed by the temperature induction. The success of prokaryotic expression is the first step of the research which studies the relationship of ubiquitin and immune in Penaeus chinesis.