猫ω型干扰素在家蚕杆状病毒表达系统的表达及活性检测  

Expression of Feline Interferon ω in Bombyx m ori Baculovirus Expression System and Determination of Its Antiviral Activity

作　　者：陈晓妍[1,2] 李皓洋 胡小元[2] 吕言娜 李轶女[2] 张志芳[2] 易咏竹[1,3] 

机构地区：[1]江苏科技大学,江苏镇江212003 [2]中国农业科学院生物技术研究所,北京100081 [3]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2015年第4期682-687,共6页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划"863"项目(No.2011AA10060-3);国家重点基础研究发展计划"973"项目(No.2012CB-114600)

摘　　要：将优化合成的猫ω型干扰素基因Fe IFN-ω2和Fe IFN-ω9克隆到杆状病毒转移载体p VL1393的多角体蛋白基因启动子下游,与orf1629基因缺损的家蚕杆状病毒Bm-Bacmid DNA共转染Bm N细胞进行同源重组,将获得的重组病毒感染家蚕5龄幼虫,利用家蚕生物反应器表达猫ω型干扰素。采用微量细胞病变抑制法在CRFK细胞/VSV*GFP系统上检测家蚕幼虫血淋巴中表达的重组猫ω型干扰素具有抗病毒活性,其中表达产物重组Fe IFN-ω2的抗病毒活性可达6.3×105U/m L,而重组Fe IFN-ω9的抗病毒活性较低。研究结果为利用家蚕生物反应器生产重组猫ω型干扰素生物制剂奠定了一定的试验基础。The optimized feline interferon ω gene Fe IFN-ω2 and Fe IFN-ω9 were artificially synthesized and cloned into baculovirus transfer vector p VL1393 to construct the recombinant plasmid under control of polyhedron gene＇s promoter.The recombinant plasmids were co-transfected into Bm N cells with orf1629-defective Bm-Bacmid DNA for homologous recombination. The obtained recombinant virus was used to infect silkworm（ Bombyx mori） larvae of the 5th instar to express feline interferon ω in silkworm bioreactor. The recombinant feline interferon ω expressed in silkworm hemolymph had antiviral activity revealed by mini-cytopathic effect inhibition assay using the CRFK cell / VSV＊ GFP system. The antiviral activity of expression products of recombinant Fe IFN-ω2 could reach 6. 3 × 10^5 U / m L,while that of recombinant Fe IFN-ω9 was low. This study provides an experimental basis for producing recombinant feline interferon ω biological agents using silkworm bioreactor.

关 键 词：猫ω型干扰素 家蚕杆状病毒表达系统 抗病毒活性 

分 类 号：Q786[生物学—分子生物学]