文献类型：期刊文献

中文题名：罗非鱼无乳链球菌DNA疫苗的构建及免疫效果研究

英文题名：Construction and Evaluation of Streptococcusagalactiae DNA Vaccine for Tilapia(Oreochromis niloticus)

作者：王蓓[1,2,3];简纪常[2,3];蔡双虎[2,3];黄郁葱[2,3];汤菊芬[2,3];吴灶和[3,4]

机构：[1]中国海洋大学水产学院;[2]广东海洋大学水产学院;[3]广东省水产经济动物病原生物学及流行病学重点实验室;[4]仲恺农业工程学院

年份：2013

期号：12

起止页码：30

中文期刊名：中国海洋大学学报：自然科学版

收录：CSTPCD、、北大核心2011、北大核心、CSCD、CSCD2013_2014

基金：国家重点基础研究发展计划项目(2009CB118704);国家重点基础研究发展计划前期项目(2011CB111600);广东省科技计划项目(2010B020309009)资助

语种：中文

中文关键词：无乳链球菌;免疫;罗非鱼;重组DNA疫苗

外文关键词：Streptococcus agalactiae immunity tilapia; recombinant DNA vaccine

中文摘要：为研究罗非鱼源无乳链球菌重组DNA疫苗对鱼体的免疫保护作用,将Sip基因克隆至真核表达载体pcDNA3.1(+)构建重组质粒pcDNA-Sip,并以肌肉注射方式免疫吉富罗非鱼(Oreochromis niloticus)。免疫后第7、28天取样,采用PCR及RT-PCR方法检测pcDNA-Sip在各个组织(包括肌肉、脾脏、头肾、鳃及肝脏)中的表达情况;分别于免疫后第7、14、21、28天采集血清,使用ELISA方法检测抗体效价;免疫28d后进行攻毒实验,计算疫苗的免疫保护率。结果表明,重组质粒pcDNA-Sip可以在上述各组织中被检测到;免疫21d后抗体效价出现峰值达1∶5 120;免疫保护率为90%。由此证实,pcDNA-Sip重组质粒可以转染到罗非鱼组织细胞,并可在鱼体中持续表达,同时有较高的免疫保护率。

外文摘要：In this paper, construction and protective immunity of the Streptococcus agalactiae recombined DNA vaccine for Oreochromis niloticus was studied. Briefly, the Sip gene of Streptococcus agalactiae was inserted into pcDNA3.1 （4-） vector and confirmed by restriction endonuclease digestion, PCR amplifica- tion and sequencing. Oreochromis niloticus was immunized with the recombined plasmid designated as pcDNA-Sip. The pcDNA-Sip expression in tissues including muscle, spleen, head kidney, gill and liver was analyzed by PCR and RT-PCR on day 7 and 28 after immunization. Antibody titer was detected with ELISA on day 7, 14, 21 and 28 after immunization. In addition, the immune protective rate of pcDNA-Sip was calculated on day 28 after immunization based on the mortality. The results indicated that pcDNA-Sip expression could be found in all studied tissues and antibody titer peaked on day 21 （1 ：5 120） after immu nization. Additionally, the relative percentage survival （RPS） value was 90% according to the mortality by S. agalactiae challenge. The recombinant plasmid pcDNA-Sip could transfect. niloticus tissues and keep expressing for a period of time, especially acquiring a high RPS.