Cloning and characterization of the 14-3-3 protein gene from the halotolerant alga Dunaliella salina.
Previous studies have demonstrated that 14-3-3 proteins exist in all the eukaryotic organisms studied; however, studies on the 14-3-3 proteins have not been involved in the halotolerant, unicellular green alga Dunaliella salina so far. In the present study, a cDNA encoding 14-3-3 protein of D. salina was cloned and sequenced by PCR and rapid amplification of cDNA end (RACE) technique based on homologous sequences of the 14-3-3 proteins found in other organisms. The cloned cDNA of 1485 bp in length had a 29.2 kDa of molecular weight and contained a 774 bp of open reading frame encoding a polypeptide of 258 amino acids. Like the other 14-3-3 proteins, the deduced amino acid sequences of the D. salina 14-3-3 protein also contained two putative phosphorylation sites within the N-terminal region (positions 62 and 67). Furthermore, an EF hand motif characteristic for Ca(2+)-binding sites was located within the C-terminal part of this polypeptide (positions 208-219). Analysis of bioinformatics revealed that the 14-3-3 protein of D. salina shared homology with that of other organisms. Real-time quantitative PCR demonstrated that expression of the 14-3-3 protein gene is cell cycle-dependent.
Wang
Tianyun
T
Laboratory for Cell Biology, Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China.
Xue
Lexun
L
Ji
Xiang
X
Li
Jie
J
Wang
Yafeng
Y
Feng
Yingcai
Y
D048948:14-3-3 Proteins / Q000737:chemistry / Q000235:genetics*; D020418:Algal Proteins / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019143:Evolution, Molecular; D020869:Gene Expression Profiling; D005796:Genes; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D019657:Solanaceae / Q000235:genetics*
D048948:14-3-3 Proteins; D020418:Algal Proteins