title: [Prokaryotic expression and antibody preparation of human  GALNT3-sol protein].
abstract: OBJECTIVE
  In order to detect the expression of GALNT3 in various tumor tissues,  the prokaryotic expression vector of human GALNT3-sol (a truncation of GALNT3 being deleted of the hydrophobic trans-membrane domain) was constructed,  and then the recombinant GALNT3-sol protein was expressed and purified from E.coli,  followed by the preparation of polyclonal antibody against GALNT3-sol and characterization of its properties.
  
  
  METHODS
  The human cDNA of GALNT3-sol (1 755 bp)was amplified from MKN45 cell line and cloned into expression vector pET5b/GALNT3-sol,  then transformed into E.coli BL21(DE3),  in which the GALNT3-sol protein was induced by IPTG and then purified by Electrophoresis.Mice were immunized with the purified protein and the anti-serum was collected at different time intervals.Properties of the anti-serum were further detected by ELISA and Western blot.
  
  
  RESULTS
  The prokaryotic expression vector of pET15b/GALNT3-sol was constructed successfully.Human GALNT3-sol protein was expressed in E.coli  after IPTG induction.The titer of the obtained anti-serum reached 1:25 600,  and its specificity was proved by Western blot.
  
  
  CONCLUSIONS
  Human GALNT3-sol protein can be successfully expressed in E.coli,  and the specific anti-human GALNT3-sol antibody can be obtained by immunization of mice,  which makes it possible to further investigate the role of GALNT3 in the progression of various tumors.
authors.affiliation: School of Life Science, Shandong University, Jinan, China. kongyu626@sohu.com
authors.forename: Yun
authors.initials: Y
authors.lastname: Kong
authors.forename: Hai-tao
authors.initials: HT
authors.lastname: Gao
authors.forename: Shu-fang
authors.initials: SF
authors.lastname: Li
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Gu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D000906:Antibodies; D017350:N-Acetylgalactosaminyltransferases; D011993:Recombinant Fusion Proteins; D000097763:Polypeptide N-acetylgalactosaminyltransferase