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目的 克隆和表达编码淋球菌IgA蛋白酶中和表位的基因。方法 通过PCR扩增淋球菌IgA蛋白酶编码基因 16 0 1～ 2 72 2位序列 ,克隆后在大肠杆菌中表达 ,表达产物用重组淋球菌IgA蛋白酶免疫血清作Western印迹。并用表达产物免疫小鼠血清与产IgA蛋白酶的标准菌株培养上清作用 ,观察免疫血清对酶活性的影响。结果 所克隆的基因在大肠杆菌中获得高效表达 ,表达产物能与重组淋球菌IgA蛋白酶免疫血清反应 ,表达产物的免疫血清能中和淋球菌IgA蛋白酶的活性。结论 所克隆的基因序列中含有淋球菌IgA蛋白酶的中和表位 ,其表达产物能中和淋球菌IgA蛋白酶的活性
Objective To clone and express the gene encoding neutralizing epitope of gonococcal IgA protease. Methods The gonococcal IgA protease encoding gene from 16 0 1 to 2 72 2 was amplified by PCR, cloned and expressed in E. coli. The expressed product was immunoblotted with immunoglobulin of gonococcus of Neisseria gonorrhoeae for Western blotting. The serum of mice was immunized with the expression product and the supernatant was cultured with a standard strain of IgA protease to observe the effect of the immune serum on the enzyme activity. Results The cloned gene was highly expressed in E. coli. The expressed product reacted with immunoglobulin gonococcal IgA protease and the immune serum of the expressed product neutralized gonococcal IgA protease activity. Conclusion The cloned gene sequence contains the Neisseria gonorrhoeae neutralization epitope of Neisseria gonorrhoeae, and its expression product can neutralize Neisseria gonorrhoeae IgA protease activity