title: [Expression and identification of mGITRL by Bac-to-Bac baculovirus expression system].
abstract: OBJECTIVE
  To express the mouse glucocorticoid-induced tumor necrosis factor receptor ligand (GITRL) protein with Bac-to-Bac baculovirus expression system.
  
  
  METHODS
  GITRL gene was obtained by double digestion using EcoR I and Sal I and cloned into the baculovirus transfer vector pFastBacHTA. Then the pFastBacHTA was transformed into competent 10BacTM E.coli cells. The transposition occurred between pFastBacHTA and bacmid and a recombinant bacmid was obtained. The positive clones were picked out and the recombinant bacmid was isolated, and then complete transfected into Tn cells for producing complete recombinant baculovirus. The baculoviral stock was amplified and the GITRL protein was expressed.
  
  
  RESULTS
  The presence of GITRL gene containing the recombinant bacmid was verified by PCR and gene sequencing. The cytopathic effect (CPE) displayed in the transfected Tn cells assumed that the transfection was successful. Western blot analysis showed that the molecular weight of mGITRL protein was about 20 KD.
  
  
  CONCLUSIONS
  The GITRL protein is expressed successfully with Bac-to-Bac baculovirus expression system, which may lay the foundation for further study on biological activity and function of GITRL protein.
authors.affiliation: Department of Immunology, School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China. zjflmj19780723@126.com
authors.forename: Jie
authors.initials: J
authors.lastname: Ma
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
authors.forename: Qing-ling
authors.initials: QL
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Xian-zhi
authors.initials: XZ
authors.lastname: Yang
authors.forename: Hua-xi
authors.initials: HX
authors.lastname: Xu
authors.forename: Sheng-jun
authors.initials: SJ
authors.lastname: Wang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D005822:Genetic Vectors; D051379:Mice; D011994:Recombinant Proteins / Q000032:analysis; D048069:Tumor Necrosis Factors / Q000235:genetics* / Q000502:physiology
chemical_list: D011994:Recombinant Proteins; C478966:Tnfsf18 protein, mouse; D048069:Tumor Necrosis Factors