【摘要】 以干酪乳杆菌为传递载体,以开发食品级安全的减蛋综合征病毒疫苗为目标,构建了一个温度敏感型自杀型质粒系统p ORZP-UKD。将该质粒电转化到干酪乳杆菌L.casei中,经过2次升温处理和5-氟尿嘧啶抗性筛选,挑选阳性克隆,采用PCR和SDS-PAGE方法进行鉴定。KnobS基因整合到干酪乳酸杆菌基因组内,并实现融合蛋白KnobS的分泌表达。表明得到了一株具有无任何选择性标记、携带有KnobS表达盒元件的基因组整合的重组干酪乳酸杆菌KnobSΔupp L.casei,且整合的外源基因能够随着细菌的复制而进行表达,为减蛋综合征病毒免疫保护性抗原KnobS疫苗的进一步研制提供了试验数据。更多还原

【Abstract】 To develop an EDSV( Egg drop syndrome virus) vaccine that live up to the standard of food safety,a temperature-sensitive suicide plasmid p ORZP-UKD was constructed using L. casei as a transmitted vector. After transforming it into L. casei using electric shock,two temperature-elevating treatments and a 5-FU screening were performed to get positive clone. Upon the identification of putative clone by PCR and SDS-PAGE,we successfully obtained a recombinant strain,KnobSΔupp L. casei,which has no selective-marker but harbors a KnobS expression box. Any exogenous gene can be integrated into this recombinant strain and express its products as bacteria replicate itself,thus providing the experimental basis for the further development of protective Knobs antigen vaccine against Egg drop syndrome virus.更多还原