title: [Cloning and prokaryotic expression of casein kinase II subunit beta gene fragment of Dirofilaria immitis].
abstract: OBJECTIVE
  To clone and express the partial fragment of Csnk2b gene of Dirofilaria immitis in prokaryotic cells, and analyze the immunoreactivity.
  
  
  METHODS
  The partial fragment of Csnk2b gene was amplified by PCR with a pair of specific primers. The PCR product was cloned into pMD18-T, and then sub-cloned to pGEX-4T-1 expression vector. The constructed plasmid pGEX-4T-1-Csnk2b was transformed into E. coli Rosetta (DE3) and followed by expression of the protein induced by IPTG. The recombinant protein was analyzed by SDS-PAGE and identified by Western blotting.
  
  
  RESULTS
  The PCR product was about 700 bp. Enzyme digestion and DNA sequencing confirmed that the recombinant plasmid pGEX-4T-1-Csnk2b was constructed. SDS-PAGE results showed that the relative molecular weight (M(r)) of the fusion protein (GST-Csnk2b) was about 45 000. GST-Csnk2b reacted positively with mouse anti-D. immitis serum.
  
  
  CONCLUSIONS
  The partial Csnk2b gene has been expressed in prokaryotic expression system and shows immunoreactivity.
authors.forename: Jun-Shan
authors.initials: JS
authors.lastname: Gao
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.forename: Hong-Lie
authors.initials: HL
authors.lastname: Hou
authors.forename: Peng-Tao
authors.initials: PT
authors.lastname: Gong
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Li
authors.forename: He
authors.initials: H
authors.lastname: Li
authors.forename: Guo-Cai
authors.initials: GC
authors.lastname: Zhang
authors.forename: Xi-Cheng
authors.initials: XC
authors.lastname: Zhang
authors.forename: Ju
authors.initials: J
authors.lastname: Yang
mesh_terms: D000818:Animals; D047390:Casein Kinase II / Q000235:genetics*; D003001:Cloning, Molecular; D004183:Dirofilaria immitis / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D047390:Casein Kinase II