| 摘要: |
| 于1996年11月–1997年5月，运用差异离心、蔗糖梯度离心、柱层析等方法从发病中国对虾分离出杆状病毒并纯化；分别用冷乙酸处理法、十二烷基硫酸钠-苯酚法提取纯化该病毒的结构蛋白与核酸、λDNA；分别用十二烷基硫酸钠煮沸法、三氯乙酸法纯化弧菌的肽葡聚糖和鸡卵粘蛋白；配制出小牛血清白蛋白、蜗牛凝集素两溶液。然后从超氧化物歧化酶（SOD）活力和溶菌活力两种免疫指标分析灭活病毒和以上提取、纯化的7种大分子物质对中国对虾的免疫效果，根据其结果筛选几种物质制成复合疫苗，测定复合疫苗对对虾的免疫作用。结果表明，病毒免疫后的SOD活力、溶菌活力均有提高。7种大分子物质免疫后，除小牛血清白蛋白外，其余6种物质的两种酶活性均有提高。以6种物质组成的复合疫苗免疫实验表明，两种酶活力提高明显（SOD活力最高峰达224.10units，溶菌活力最高峰为0.122units）。说明复合疫苗具有提高SOD和溶菌活力的作用，可以增强中国对虾的防御能力。 |
| 关键词: 中国对虾 病毒 弧菌 糖蛋白 免疫激活剂 复合疫苗 |
| DOI： |
| 分类号: |
| 基金项目:国家攀登计划B资助项目，PDB6－6－3号;国家“九五”科技攻关项目，055970311号 |
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| PRIMARY STUDIES ON COMPLEX VACCINE OF PENAEUS CHINENSIS |
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DING Yu1, WANG Lei1, LI Guang-you2
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1.Institute of Oceanology, The Chinese Academy of Sciences, Qingdao, 266071;2.First Institute of Oceanography, State Oceanic Administration, Qingdao, 266003
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| Abstract: |
| From November 1996 to may 1997, rod viruses of P. chinensis were separated and purifies from disease prawns P. chinensis through differential centrifugation, sucrose gradient centrifugation and column chromatograph. Their capsid protein was purified using ice-acetic acid method, an SDS-phenol method was applied to extract their DNA as well as λDNA. Peptidoglycan of Vibrio alginolyticus (by SDS-boiling method), and chicken ovomucin were also extracted by chloroacetic acid method. At last buffalo serum albumen and snail agglutinin solution were prepared with a phosphate buffer. The effects of all above materials including inactivated viruses and seven kinds of macromolecular compounds on the immune potency of P. chinensis after injected into prawns were determined by SOD and bacteriolytic activities. According to the above results several compounds were selected to form complex vaccine, whose immune potency was then evaluated. The results show that SOD and bacteriolytic activities of the test group were much more intensive than the control group after the viruses were injected. Six of the seven compounds except buffalo serum albumen can activate two enzymes activities; the SOD activity of the test group can reach 224.10 units. but that of the control group was only between 58.25–72.21 units. The bacteriolytic activity of the test group would reach 0.132 units, but that of the control group was only between 0.019 and 0.028 units. Further, the two enzymes activities of the test group were much higher than the control after the complex vaccine containing six types of materials were inoculated. In the test group maximum of SOD activity could reach 222.90 units, and that of bacteriolytic activity was 0.122 units. However, in the control group the SOD activity was changing from 78.43 to 87.86 units and the bacteriolytic activity was from 0.025 to 0.041 units. It is suggested that this complex vaccine can strengthen SOD and bacteriolytic activities and can increase immune potency of P. chinensis. |
| Key words: Penaeus chinensis, Virus, Vibrio, Glycoprotein, Immune stimulant, Complex vaccine |
