【摘要】 目的:研究弓形虫表面抗原p22基因与p30基因的联合表达。方法:用RT-PCR及PCR方法获得p22和p30基因,联合构建在表达载体中,经酶切与测序鉴定后,用IPTG对工程菌进行诱导表达,表达产物行SDS-PAGE蛋白凝胶电泳及Western-blot免疫印迹检测。结果:经RT-PCR可得到长约438bp的p22外显子基因片段,蛋白电泳结果显示,阳性重组菌在66.5Kda位置上明显多一条带,此条带可与p22抗体结合并使免疫印迹显示阳性结果。结论:弓形虫表面抗原p22基因的有效基因片段与p30基因片段联合表达后,在融合蛋白中仍具有免疫原性。更多还原

【Abstract】 Objective:To obtain the exon of the p22gene,check up the activity of the ex-pression products,then link it with the p30gene,and identify its activity in the fusion protein.Meth ods:The recombinant gene of p22and p30of toxoplasma was obtained through the RT-PCR and common PCR.After identification with enzyme cutting and sequencing,the expression of engineer-ing bacteria were induced by IPTG.The expression products were identified by SDS-PAGE and Western-Blot analysis.Re sults:The exon of p22was about 438bp.The positive recombinant bacteria had an extra protein band at66.5Kda site compared with the negative control.The Western-Blot with the p22antibody showed there was a brown band at the same site.Conclusion:The membrane anti-gen p22gene with the intron can be abstracted with the RT-PCR,and expression of p22-p30fusion protein was successfully induced.更多还原