Molecular cloning and expression analysis of stress-inducible protein I gene of Miichthys miiuy.

Abstract

The full length of stress-inducible protein Ⅰ(STI1) gene cDNA was cloned from Miichthys miiuy based on the obtained EST sequence using rapid amplification of cDNA ends polymerase chain reaction(RACE-PCR).The STI1 gene cDNA consisted of 2 194 nucleotides including 5' untranslated regions(UTR) of 31 nucleotides,3' UTR of 634 nucleotides and open reading frame with 1 629 nucleotides encoding 542 amino acid residues.The molecular structure of STI1 consisted of five potential casein kinase Ⅱ phosphorylation sites,one polyproline stretch,two nuclear localization signals and nine tetratricopeptide repeats(TPR).The amino acid sequence of the STI1 gene shared 76.5%-94.3% identity with those of previously reported other species STI1 gene including conserved eight cysteines.Phylogenetic tree analysis based on amino acid sequences of STI1 gene generated by neighbor-joining method suggested that four teleosts clustered together,and were close genetic relationship between M.miiuy and Pseudosciaena crocea.Real-time PCR showed that the STI1 gene was mainly detected in muscle and kidney.