title: Expression and purification of half-smooth tongue sole (Cynoglossus semilaevis) CSDAZL protein.
abstract: The csdazl gene is a sex related gene of half-smooth tongue sole (Cynoglossus semilaevis). Our research group have cloned full length cDNA of csdazl, and studied its expression pattern. To get the further information of csdazl, we constructed a prokaryotic expression plasmid, pET-32a-CSDAZL, expressed in Escherichia coli BL21 and purified the fusion protein by His Trap. In order to detect the biological activity of the fusion protein, we injected the protein with liposome into fish, and detected other sex-related genes' mRNA expression. The results showed that the expression levels of half-smooth tongue sole female-related genes Cyp19a and Foxl2 significantly decreased between 6 and 24 h; however, both genes' expressions returned to their normal levels 72 h after injection, indicating that recombinant CSDAZL protein could down-regulated the expression of female-related genes, Foxl2 and Cyp19a genes, implying that the fusion protein has biological activity and csdazl plays a role in sex differentiation by regulating sex related genes' expression.
authors: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
authors: Kailin
authors: K
authors: Wang
authors: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
authors: Hong
authors: H
authors: Zhang
authors: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
authors: Qiaomu
authors: Q
authors: Hu
authors: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
authors: Changwei
authors: C
authors: Shao
authors: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China. Electronic address: chensl@ysfri.ac.cn.
authors: Songlin
authors: S
authors: Chen
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D029941:Fish Proteins / Q000008:administration & dosage / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005412:Flatfishes / Q000235:genetics*; D005786:Gene Expression Regulation; D008297:Male; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D019849:Sex Determination Processes; D014162:Transfection
chemical_list: D029941:Fish Proteins; D011993:Recombinant Fusion Proteins
keywords: CSDAZL; Cynoglossus semilaevis; Fusion protein; Protein transfection