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作 者:丁农 张金卫 李江涛 鱼南洋 冯世民 沈建华 徐森华
出 处:《安徽农业科学》2016年第1期220-222,共3页Journal of Anhui Agricultural Sciences
基 金:浙江省公益技术农业项目(2014C32054);湖州市公益技术应用研究项目(2013GY09)
摘 要:[目的] 研究家蚕生物反应器规模化共表达基因工程产品猪α+γ干扰素的关键技术,为家蚕生物反应器规模化共表达猪α+γ干扰素的生产提供技术支持。[方法]采用α干扰素ELISA检测试剂盒检测蚕品种、接种方式、蚕蛹冷藏对共表达干扰素重组杆状病毒rBmNPV(BmIFNα+γ)表达量的影响。 [结果]不同蚕品种对重组杆状病毒的表达效率差异较大,春用品种选择皓月×菁松,夏秋用品种选择白玉×秋丰。机械注射接种效率比手工注射提高了3倍以上,检测产品表达量相差不大。蚕在上蔟后25 ℃保护,第5~9天是注射适期,以第6~8天为最佳。蚕茧冷藏于4~5 ℃ 10 d以内,对重组杆状病毒在蚕蛹中的表达效率影响不大。[结论]采用高表达效率的蚕品种以及结合机械的使用和规范操作可以确保规模化生产的成功。[ Objective ] To study the key technologies of silkworm bioreactor large scale porcine co-expression α+γ interferon, and provide technical support for the production of silkworm bioreactor large scale porcine co-expression α+γ interferon. [ Method]or interferon ELISA testing kits were used for testing the effects of silkworm varieties, inoculation methods and silkworm pupae cold storage on the amount of expression of co-expression interferon recombinant baculovirus rBmNPV (BmIFN-α+γ). [ Result] Different kinds of silkworms differ largely in expression efficiency to recombinant baculovirus, with Haoyue x Jingsong higher and Qiufeng × Baiyu lower. Mechanical injection inoculation was over three times more efficient than manual, with similar expression of tested products. After mounting, the silkworms needed to be protected at a temperature of 25℃. The fifth to ninety day was the optimum time for injection, with the sixth to eighth the best. Cocoons were refrigerated at 4-5℃ for ten days, which had a little influence on the expression efficiency of recombinant baculovirus in cocoons. [ Conclusion] The use of high expression efficiency silkworm varieties and the standardized use of machines can ensure the success of large scale production. Key words Silkworm bioreactor; Co-expression ;Porcine ;α+γ Interferon
分 类 号:S882.31[农业科学—特种经济动物饲养]
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