title: [Cloning, prokaryotic expression of CML66 and preparation of its polyclonal antibody].
abstract: OBJECTIVE
  To clone and express CML66 cDNA and to prepare rabbit anti-CML66 antibody.
  
  
  METHODS
  cDNA isolated from the testis using RT-PCR was cloned into pGEMT. After sequencing, the cDNA was inserted into prokaryotic expression vector pET32b(+). The recombinant vector was transformed into BL-21(DE3) through electroporation. 6xHis-tagged CML66 expression was then induced by IPTG. The protein was purified through Ni(2+) affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified protein was injected into rabbits to prepare polyclonal antibody.
  
  
  RESULTS
  The cloned cDNA sequence was identical with that previously reported. The target protein was successfully purified. And rabbit's anti-serum with high titer was obtained.
  
  
  CONCLUSIONS
  We have cloned CML66 successfully, expressed and purified the protein in E.coli.Furthermore,rabbit polyclonal antibody has been obtained.
authors.affiliation: Shanghai Institute of Immunology, Shanghai Second Medical University, Shanghai 200025, China. youqiang@etang.com
authors.forename: Qiang
authors.initials: Q
authors.lastname: You
authors.forename: Lin-ling
authors.initials: LL
authors.lastname: Cheng
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Shu-jun
authors.initials: SJ
authors.lastname: Wang
authors.forename: Hui-zhen
authors.initials: HZ
authors.lastname: Zhang
authors.forename: Hai-liang
authors.initials: HL
authors.lastname: Ge
mesh_terms: D000906:Antibodies / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008970:Molecular Weight
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D018076:DNA, Complementary; C431278:NUDCD1 protein, human