利用改构的pEGFP-N1-ie1强启动子载体构建含vp28基因的核酸疫苗,通过肌肉注射和饲料添加的形式免疫对虾,分析各实验组相对保护率和免疫基因(Dicer,Argonaute,STAT和Lyzome)表达,研究构建vp28重组核酸疫苗对凡纳滨对虾的保护效果.RT-PCR结果表明,注射和口服vp28组均可在对虾鳃组织中检测到vp28基因表达.在注射组中vp28的相对保护率为22.4％,饲料免疫组中vp28的相对保护率可达到36.84％.相对于对照组,vp28的注射组的STAT基因和Dicer基因表达量于第7天达到最大值.Argonaute基因表达量于第3天达到最大值,以后呈下降趋势.溶菌酶基因表达量一直呈相对较高的趋势.vp28的饲料投喂组的STAT基因和Argo-naute基因表达量于第7天达到最大值,以后呈下降趋势.Dicer基因表达量第3天达到最大值.实验结果表明,构建的vp28核酸疫苗在对虾抗WSSV防治中具有潜在应用价值.%In this study,pEGFP-N1-ie1 was reconstructed by inserting WSSV immediate early gene,and vp28 was cloned in a new constructed vector.The survival rates and antiviral associated factors (Dicer,Argonaute,STAT and Lyzome) were detected in Litopenaeus van-namei after immunity by injection and oral-feeding.RT-PCR results showed that vp28 was detected in gill of shrimp by injection and oral-feeding immunity treatment.The relative protection rate of vp28 group reached 22.4% (injection group) and 36.84% (oral feeding group).The STAT and dicer mRNA expression in vp28 group were significantly higher than that of control group after injection treatment for 7 d.The argonaute mRNA expression in vp28 group was significantly higher than that of control group after injection treatment for 3 d and then decreased.The lysozyme mRNA expression in vp28 injection group was consistently significantly higher than that of control group.But the STAT and Argonaute mRNA expression in vp28 group was significantly higher than that of control group on the 7 th d of oral-feeding treatment and then decreased.The dicer mRNA expression in vp28 group was significantly higher than that of control group on the 3rd day of oral treatment.These results demonstrated that vp28 DNA vaccine has a potential utility for control of WSSV disease in shrimp.
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