title: [Prokaryotic Expression and Immunoreactivity Analysis of α-8 Giardin in Giardia lamblia].
abstract: OBJECTIVE
  To clone and express α8-giardin gene of Giardia lamblia, and analyze its immunoreactivity.
  
  
  METHODS
  The open reading frame (ORF) of α8-giardin gene was amplified by PCR. The PCR product was cloned into prokaryotic expression vector pET-30a (+) with restriction enzymes EcoR I and Xho I . The recombinant vector pET30a (+)-α8-giardin was transformed into E. coli BL21 (DE3), and the positive clones were then selected. The constructed pET30 a (+)-α8-giardin was induced with IPTG for expression, and purified through Ni-affinity chromatography. The recombinant protein was examined by SDS-PAGE and Western blotting.
  
  
  RESULTS
  The length of α8-giardin gene was 930 bp. PCR and restriction enzyme digestion analysis confirmed the construction of recombinant plasmid pET30a (+)-α8-giardin. SDS-PAGE and Western blotting analysis showed that the recombinant protein rGiardin (about M, 36,000) was expressed in E. coli as inclusion body protein, and reacted positively with anti-His tag antibody and rabbit anti-G. lamblia serum.
  
  
  CONCLUSIONS
  The recombinant plasmid pET30a (+)-α8-giardin is constructed, and the purified rGiardin protein shows immunoreactivity.
authors.forename: Chao-jun
authors.initials: CJ
authors.lastname: Wei
authors.forename: Ling
authors.initials: L
authors.lastname: Wu
authors.forename: Qin
authors.initials: Q
authors.lastname: Wei
authors.forename: Yan-juan
authors.initials: YJ
authors.lastname: Jia
authors.forename: Si-qi
authors.initials: SQ
authors.lastname: Lu
mesh_terms: D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005822:Genetic Vectors; D016829:Giardia lamblia; D016133:Polymerase Chain Reaction; D011506:Proteins; D011994:Recombinant Proteins
chemical_list: D000906:Antibodies; D011506:Proteins; D011994:Recombinant Proteins