犬α干扰素在家蚕杆状病毒表达系统中的表达及其抗病毒活性的测定

doi:10.13560/j.cnki.biotech.bull.1985.2015.06.022

生物技术通报 ›› 2015, Vol. 31 ›› Issue (6): 216-220.doi: 10.13560/j.cnki.biotech.bull.1985.2015.06.022

• 研究报告 • 上一篇

犬α干扰素在家蚕杆状病毒表达系统中的表达及其抗病毒活性的测定

李皓洋¹ 胡小元¹ 易咏竹² 杨鑫¹ 张志芳¹ 李轶女¹

1.中国农业科学院生物技术研究所，北京100081；2.中国农业科学院蚕业研究所，镇江 212018

收稿日期:2015-03-17 出版日期:2015-06-19 发布日期:2015-06-20
作者简介:李皓洋，男，硕士，研究方向：昆虫杆状病毒表达系统；E-mail：815579279@qq.com
基金资助:
国家高技术研究发展计划“863”计划(2011AA100603)，国家重点基础研究发展计划“973”计划(2012CB114600)

Expression of Canine Interferon Alpha in Silkworm-baculovirus Expression System and the Antiviral Activity Assay

Li Haoyang¹, Hu Xiaoyuan¹, Yi Yongzhu², Yang Xin¹, Zhang Zhifang¹, Li Yinü¹

1. Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081；2. The Sericultural Research Institute，Chinese Academy of Agricultural Sciences，Zhenjiang 212018

Received:2015-03-17 Published:2015-06-19 Online:2015-06-20

摘要/Abstract

摘要： 犬α干扰素(CaIFNα)在犬病防治过程中应用广泛。旨在开发一种高效的CaIFNα表达方法。首先按家蚕密码子的偏好性对GenBank中的一个CaIFNα基因进行了优化与合成，将其克隆到杆状病毒转移载体pVL1393中，并与亲本病毒BmBacmid共转染家蚕细胞进行细胞内重组。得到的重组病毒用于感染家蚕幼虫，收集发病幼虫的蚕血淋巴用于CaIFNα检测。采用细胞病变抑制法在MDCK-VSV*GFP系统中测定其抗病毒活性。结果显示家蚕表达的CaIFNα能有效抑制VSV*GFP在细胞内的复制，其抗病毒活性不低于1.78×10⁶U/mL。

关键词: 犬α干扰素, 家蚕杆状病毒表达系统, 抗病毒活性检测

Abstract: Canine interferon alpha(CaIFNα)was widely used in the prophylaxis and cure of canine diseases. The purpose of this study is to develop an efficient method to express CaIFNα. A canine interferon alpha gene in GenBank was optimized according to the codon bias of silkworm and synthesized, and then cloned into the baculovirus transfer vector pVL1393 to construct the recombinant plasmid pVL-CaIFNα. The pVL-CaIFNα was co-transfected with the BmBacmid DNA into silkworm cells and after in vivo recombination. The recombinant virus was gathered and used to infect silkworm larvae. The hemolymph of infected larvae was collected for antiviral activity assay. The antiviral activity of expressed CaIFNα was examined on Madin-Darby canine kidney cells infected with the vesicular stomatitis virus expressing green fluorescent protein(VSV*GFP). The result showed that CaIFNα expressed in silkworm can inhibit the multiplication of VSV*GFP and the antiviral activity was about 1.78×10⁶U/mL.

Key words: canine interferon alpha, silkworm-baculovirus expression system, antiviral activity assay

李皓洋, 胡小元, 易咏竹, 杨鑫, 张志芳, 李轶女. 犬α干扰素在家蚕杆状病毒表达系统中的表达及其抗病毒活性的测定[J]. 生物技术通报, 2015, 31(6): 216-220.

Li Haoyang, Hu Xiaoyuan, Yi Yongzhu, Yang Xin, Zhang Zhifang, Li Yinü. Expression of Canine Interferon Alpha in Silkworm-baculovirus Expression System and the Antiviral Activity Assay[J]. Biotechnology Bulletin, 2015, 31(6): 216-220.

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犬α干扰素在家蚕杆状病毒表达系统中的表达及其抗病毒活性的测定

Expression of Canine Interferon Alpha in Silkworm-baculovirus Expression System and the Antiviral Activity Assay

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