关键词: 　蓝氏贾第鞭毛虫；&alpha, 8-贾第素；克隆；原核表达

Abstract:

【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 EcoRⅠ and XhoⅠ. The recombinant vector pET30a (+)-α8-giardin was transformed into E. coli BL21(DE3), and the positive clones were then selected. The constructed pET30a(+)-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 Mr 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.  Conclusion The recombinant plasmid pET30a(+)-α8-giardin is constructed, and the purified rGiardin protein shows immu-noreactivity.

Key words: Giardia lamblia, α8-giardin, Clone, Prokaryotic expression