CAJ | 학술논문

为了原核表达猪源马链球菌兽疫亚种去除信号肽的类M蛋白质,并分析该蛋白质对小鼠的免疫保护力,克隆了马链球菌兽疫亚种CY菌株去除信号肽的类M蛋白质基因Szp,并将Szp基因插入pET28a表达载体, IPTG诱导,获得重组类M蛋白质,弗氏佐剂乳化重组类M蛋白质后,3次免疫小鼠,用同源菌株CY攻击。诱导获得分子量约5.8×104的重组蛋白质,免疫印迹结果表明该重组类M蛋白质能够被马链球菌兽疫亚种猪多抗识别。类M蛋白质免疫小鼠的存活率为60％,弗氏佐剂乳化灭活CY免疫小鼠的存活率为70％,PBS对照组小鼠攻毒后6 d内全部死亡。表达的类M蛋白质可以给予ICR小鼠部分的免疫保护力,保护效果略低于CY全菌灭活苗,提示可以对马链球菌兽疫亚种多种免疫保护性抗原进行联合,从而进一步提高对动物的免疫保护力。
위료원핵표체저원마련구균수역아충거제신호태적류M단백질,병분석해단백질대소서적면역보호력,극륭료마련구균수역아충CY균주거제신호태적류M단백질기인Szp,병장Szp기인삽입pET28a표체재체, IPTG유도,획득중조류M단백질,불씨좌제유화중조류M단백질후,3차면역소서,용동원균주CY공격。유도획득분자량약5.8×104적중조단백질,면역인적결과표명해중조류M단백질능구피마련구균수역아충저다항식별。류M단백질면역소서적존활솔위60％,불씨좌제유화멸활CY면역소서적존활솔위70％,PBS대조조소서공독후6 d내전부사망。표체적류M단백질가이급여ICR소서부분적면역보호력,보호효과략저우CY전균멸활묘,제시가이대마련구균수역아충다충면역보호성항원진행연합,종이진일보제고대동물적면역보호력。
An open reading frame encoding M-like protein from strain CY of Streptococcus equi subsp. zooepidemicus of porcine origin without signal sequence was amplified by PCR using chromosomal DNA of CY as template. Amplicon was inserted into plasmid pET28a and transformed into Escherichia coli BL21 (DE3). A His-tag fusion protein SzP was then in-duced with IPTG. Mice were immunized three times with SzP and challenged by the homologous strain CY. The fusion pro-tein SzP with molecular weight about 5. 8×104 showed the specific immunoreactivity with pig antiserum against strain CY. Six SzP-vaccinated mice survied out of 10 tested mice, and the figure was 7 for whole cell bacterin-vaccinated mice. Meanwhile, the control mice vaccinated with PBS all died 6 days post-infection. It is indicated that the fusion protein SzP delivered certain protection against Streptococcus equi subsp. zooepidemicus.