title: [Preparation of monoclonal antibody against phosphinothricin acetyltransferase].
abstract: OBJECTIVE
  To express phosphinothricin acetyltransferase (PAT) with biological activity and prepare monoclonal antibodies against PAT.
  
  
  METHODS
  The full length bar gene was cloned by PCR and inserted into prokaryotic expression vector pET28a⁺. The recombinant plasmid pET28-bar was transformed into E.coli BL21(DE3), and under the induction of IPTG, PAT was expressed. The expressed protein was purified by Ni⁺; affinity chromatography to analyze its activity. The purified PAT was used to immunize BALB/c mice, and then the spleen cells from the immunized mice were fused with Sp2/0 cells. The hybridoma clones secreting antibodies against PAT were isolated by indirect ELISA and then subcloned.
  
  
  RESULTS
  Soluble PAT was expressed in E.coli. The purified PAT had the activity of acetyltransferase. We totally prepared 9 hybridoma cell lines which secreted specific anti-PAT monoclonal antibodies.
  
  
  CONCLUSIONS
  The expressed recombinant PAT can be used for biological reagent to prevent and relieve herbicide damage. Monoclonal antibodies against PAT may be used to detect the transgenic products.
authors: College of Horticulture, Northeast Agricultural University, Harbin 150030, China. eo_777654@sina.com
authors: Xudong
authors: X
authors: Gao
authors: Yongzhi
authors: Y
authors: Wang
authors: Shengfeng
authors: S
authors: Shi
authors: Zhongpeng
authors: Z
authors: Li
authors: Xiaoyu
authors: X
authors: Li
authors: Wenjing
authors: W
authors: Xu
authors: Zhengkun
authors: Z
authors: Zhang
authors: Yang
authors: Y
authors: Lu
authors: Jiashi
authors: J
authors: Zhang
authors: Qiyun
authors: Q
authors: Li
authors: Jingang
authors: J
authors: Wang
mesh_terms: D000123:Acetyltransferases / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology; D003001:Cloning, Molecular; D004789:Enzyme Activation; D015870:Gene Expression; D006825:Hybridomas; D051379:Mice; D011993:Recombinant Fusion Proteins
chemical_list: D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase