转基因聚球藻7942中vp28基因表达效率及其光合特性分析  

vp28 Gene Expression and Photosynthetic Characteristics of Transgenic Synechococcus sp. PCC 7942

作　　者：庄旻敏 贾晓会 施定基[3] 朱嘉诚 冯思豫 何培民 贾睿 ZHUANG Min-min1,JIA Xiao-hui1,2,SHI Ding-ji3,ZHU Jia-cheng1,FENG Si-yu1,HE Pei-min1,JIA Rui1(1 School of Ocean Ecology and Environment Shanghai Ocean University, Shanghai 201306 China;2 School of Biosciences,Beijing Normal University,Beijing 100875, China;3 Institute of Botany, Chinese Academy of Sciences, Beijing 100093, Chin)

机构地区：[1]上海海洋大学海洋生态与环境学院,上海201306 [2]北京师范大学生命科学学院,北京100875 [3]中国科学院植物研究所,北京100093

出　　处：《中国生物工程杂志》2018年第4期30-37,共8页China Biotechnology

基　　金：国家863计划(2014AA093506);上海市科技兴农推广项目(沪农科推字(2017)第1-13号);上海市科委项目(16391903500,17391902200)资助项目

摘　　要：背景：对虾白斑综合征病毒（white spot syndrome virus,WSSV）是对虾养殖业中危害最严重的病毒之一,至今尚无规模应用的有效药物防治方法。但近年来在WSSV免疫防治上进展较大。Vp28蛋白是WSSV囊膜上的主要结构蛋白,2004年以来其编码基因已在8种宿主中表达成功,在实验室试验中对WSSV的防治疗效显著,但目前尚未见到其在对虾产业中的应用。目的：利用对虾的天然饵料聚球藻表达Vp28重组蛋白,这种药食同源可简化操作,降低成本,有助其在生产中应用。方法：用荧光定量PCR方法检测转vp28基因聚球藻7942中vp28基因的表达效率。通过氧电极的方法测得转vp28基因型聚球藻在不同温度、光照、pH和盐度下的光合活性变化,找到它的最适生长条件。结果：检测了vp28基因表达效率为9.52%,是在鱼腥藻7120表达效率的3倍。最适采收时间是对数生长后期（15d左右）。转基因型蓝藻7 942的最适生长条件是：温度为40℃,盐度为0～0.1mol/L NaCl,pH为7.5,光强为450μmol/（m^2·s）。结论：确定了vp28基因在聚球藻中的表达效率及该转基因藻的最适培养条件,这些研究结果为用转vp28基因型聚球藻7942规模制备药食同源的口服剂提供了依据。White spot syndrome virus（ WSSV） is one of the most harmful viruses in the shrimp industry,and there has been no effective drug on mass scale up to now. In recent years,the immunological prevention of WSSV from shrimp has been progressed hopefully. Vp28 protein is the major structural component on the envelope of WSSV. Since 2004,its coding genes have been expressed in 8 species,and the control of WSSV has been proved remarkably in the laboratory. However,it has not been applied in shrimp industry yet. The shrimp bait,Synechococcus sp. PCC7942 was used as the acceptor to express vp28 gene,and this homology of medicine and food may be helpful for its application in the shrimp industry. The expression efficiency of vp28 in transgenic Synechococcus sp. PCC7942 has been detected by RT-q PCR method. And photosynthetic characteristics of transgenic Synechococcus sp. PCC7942 at different temperature,illumination,pH and salinity have been measured by the method of oxygen electrode. The expression efficiency of vp28 gene was 9. 52% which was three times higher than that of Anabaena sp. PCC 7120. The most suitable harvest time was in late logarithm growth（ the15 thd）. The optimum growth conditions of transgenic Synechococcus sp. PCC7942 were as follows： the temperature 40℃,the salinity 0 ～ 0. 1 mol/L NaCl,pH 7. 5,light intensity 450μmol/（ m^2·s）. These data may be useful for scale preparation of oral drug made from transgenic Synechococcus sp. PCC7942.

关 键 词：转vp28基因聚球藻 光合活性 实时荧光定量PCR vp28基因表达效率 

分 类 号：Q789[生物学—分子生物学]