title: [Expression, purification and identification of fusion protein mTSLPR-Ig].
abstract: OBJECTIVE
  To construct an adenovirus vector (Ad-mTSLPR-Ig) expressing the fusion protein of the extracellular domain of mouse TSLPR and the Fc fragment of mouse Ig, further purify and assess the soluble fusion protein mTSLPR-Ig.
  
  
  METHODS
  The gene of mTSLPR extracellular domain was amplified from cDNA library of mouse thymus by PCR. Then an adenovirus vector (Ad-mTSLPR-Ig) expressing the fusion protein of mTSLPR and the Fc fragment of mouse Ig was constructed. After Ad-mTSLPR-Ig was transfected into COS-7 cells, the supernatants were collected. The fusion proteins in the supernatants were detected by double antibody sandwich ELISA. The soluble fusion proteins mTSLPR-Ig were purified by protein A affinity chromatography.
  
  
  RESULTS
  The gene sequence of Ad-mTSLPR-Ig was confirmed by DNA sequencing, and the recombinat adenoviruses harboring mTSLPR-Ig were successfully obtained after the infection of Ad-mTSLPR-Ig. Fusion proteins mTSLPR-Ig in the supernatants were detected by ELISA assay. The purified fusion protein were obtained by affinity chromatography, and identified by Western blot.
  
  
  CONCLUSIONS
  The soluble fusion proteins mTSLPR-Ig have been obtained successfully, which enables further study of its biological activity.
authors.affiliation: Nanjing General Hospital of Nanjing Military Region, Department of Respiratory Medicine, Nanjing 210002, China. Zhangfanglab@yahoo.com.cn
authors.forename: Fang
authors.initials: F
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
authors.forename: Gang
authors.initials: G
authors.lastname: Huang
authors.forename: Zi-ling
authors.initials: ZL
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Hu
authors.forename: Yong
authors.initials: Y
authors.lastname: Song
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005822:Genetic Vectors / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D007136:Immunoglobulins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D007141:Immunoglobulin Fc Fragments; D007136:Immunoglobulins; D018121:Receptors, Cytokine; D011993:Recombinant Fusion Proteins; C410591:Tslpr protein, mouse