title: [Expression and purification of two kinds of alternative splicing mouse Era].
abstract: OBJECTIVE
  To express and purify two alternative splicing mouse Era proteins and detect whether anti-human Era antibody can be used in the study of mouse Era proteins.
  
  
  METHODS
  Two fusion protein expression vectors, pMAL-meraW and pMAL-meraS, were constructed, then the MBP-mEra proteins were expressed in E. coli. The target proteins were purified by amylose affinity chromatography. The specificity of rabbit anti-human Era antibody to the proteins was identified by Western blot.
  
  
  RESULTS
  The expressed MBP-mEraW and MBP-mEraS proteins constituted approximately 17% and 19% of the total bacterial proteins. The purity of the fused proteins was 67% and 61% respectively after amylose affinity chromatography. Rabbit anti-human Era antibody had high specificity to these two kinds of splicing mouse Era proteins.
  
  
  CONCLUSIONS
  Two fusion mera genes could be expressed in E. coli by using gene recombination technique. The high specificity of rabbit anti-human Era antibody to the two splicing mouse ERA proteins indicates that this antibody can be used to study the function of these two kinds of splicing mouse Era.
authors.affiliation: Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Dong
authors.forename: Su-min
authors.initials: SM
authors.lastname: Chen
authors.forename: Zong-ling
authors.initials: ZL
authors.lastname: Ji
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
authors.forename: Nan-chun
authors.initials: NC
authors.lastname: Chen
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D019204:GTP-Binding Proteins; D015870:Gene Expression; D005818:Genetic Engineering; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: C540137:Era protein, mouse; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D019204:GTP-Binding Proteins