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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。
The nucleic acid vaccine containing vp28 gene was constructed by using the modified pEGFP-N1-ie1 strong promoter vector. The shrimp were immunized by intramuscular injection and feedstuff, and the relative protection rates and immune genes (Dicer, Argonaute, STAT and Lyzome ) Expression, to study the construction of vp28 recombinant nucleic acid vaccine on the protection of Litopenaeus vannamei. RT-PCR results showed that vp28 gene expression was detected in shrimp gill tissues both in injected and oral vp28 groups. The relative protection rate of vp28 in the injection group was 22.4%, the relative protection rate of vp28 in the feed immunization group could reach 36.84%. Compared with the control group, the expression level of STAT gene and Dicer gene in the injection group of vp28 reached the maximum on the 7th day. Argonaute gene expression reached its maximum on the third day and then decreased. Lysozyme gene expression has been a relatively high trend. The expression of STAT gene and Argo-naute gene in the feed-fed group of vp28 reached the maximum on the 7th day and then decreased. Dicer gene expression reached the maximum on the 3rd day. The experimental results show that the constructed vp28 nucleic acid vaccine has potential application in the prevention and control of WSSV in shrimp.