|本期目录/Table of Contents|

[1]梅文枫,钱月忠,史翠萍,等.基于RD BmBacmid载体构建表达GFP蛋白的重组杆状病毒的方法[J].浙江理工大学学报,2014,31-32(自科6):672-677.
 MEI Wen feng,QIAN Yue zhong,SHI Cui ping,et al.A Method of Constructing Recombinant Baculovirus Expressing GFP Protein Based on RD BmBacmid Carrier[J].Journal of Zhejiang Sci-Tech University,2014,31-32(自科6):672-677.
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基于RD BmBacmid载体构建表达GFP蛋白的重组杆状病毒的方法()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第31-32卷
期数:
2014年自科6期
页码:
672-677
栏目:
(自科)生物与生命科学
出版日期:
2014-11-10

文章信息/Info

Title:
A Method of Constructing Recombinant Baculovirus Expressing GFP Protein Based on RD BmBacmid Carrier
文章编号:
1673-3851 (2014) 06-0672-06
作者:
梅文枫 钱月忠 史翠萍 陈健
浙江理工大学 a. 生物化学研究所; b. 浙江省家蚕生物反应器和生物医药重点实验室, 杭州 310018
Author(s):
MEI Wenfeng QIAN Yuezhong SHI Cuiping CHEN Jian
a. Institute of Biochemistry; b. Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, Zhejiang Sci-Tech University, Hangzhou 310018, China
关键词:
杆状病毒 RD BmBacmid PCR 共转染 绿色荧光蛋白
分类号:
Q812
文献标志码:
A
摘要:
研究构建一种快速表达GFP蛋白的重组杆状病毒的方法。在复制缺陷型的家蚕杆状病毒(BmNPV)穿梭载体RD BmBacmid基础上,利用PCR方法合成两端分别含有50 bp左右同源臂的绿色荧光蛋白基因(AcGFP)片段和线性化的RD BmBacmid共转染BmN细胞或家蚕幼虫,可以一步获得重组病毒,表达绿色荧光蛋白。实验结果表明,含有AcGFP基因的重组杆状病毒构建成功,在BmN细胞和家蚕幼虫中分别表达了绿色荧光蛋白。因而,本研究成功建立了一种快速构建重组杆状病毒并能够表达GFP蛋白的新方法。

参考文献/References:

[1] Fernandez J M, Hoeffler J P. Gene Expression Systems: Using Nature for the Art of Expression[M]. NewYork: Academic Press, 1998: 331-359.
[2] Maeda S, Kawai T, Obinata M, et al. Production of human αinterferon in silkworm using a baculovirus vector[J]. Nature, 1985, 315(6020): 592-594.
[3] Hitchman R B, Possee R D, King L A. Baculovirus expression systems for recombinant protein production in insect cells[J]. Recent Patents on Biotechnology, 2009, 3(1): 46-54.
[4] 韦永龙, 李轶女, 张志芳, 等. 杆状病毒表达系统及其应用进展[J]. 生物技术通报, 2010, 10: 1-7.
[5] Wu X, Cao C, Xu Y, et al. Construction of a host rangeexpanded hybrid baculovirus of BmNPV and AcNPV, and knockout of cysteinase gene for more efficient expression[J]. Science in China Series C: Life Sciences, 2004, 47(5): 406-415.
[6] King L A, Possee R D. The Baculovirus Expression System: a Laboratory Guide[M]. London: Chapman & Hall London, 1992: 127-140.
[7] Wu X, Yang G, Hu J. A recombinant rescue linearizable BmNPV baculovirus BmBacPAK: China, 98110962.2[P]. 199807-17.
[8] Motohashi T, Shimojima T, Fukagawa T, et al. Efficient largescale protein production of larvae and pupae of silkworm by Bombyx mori nuclear polyhedrosis virus bacmid system[J]. Biochem Biophys Res Commun, 2005, 326(3): 564-569.
[9] 费伟强, 陈琴, 陈倩, 等. 复制缺陷型 BmNPV 载体的构建及初步应用[J]. 蚕业科学, 2013, 39(3): 0514-0521.
[10] Costantino N. Enhanced levels of λ Red mediated recombinants in mismatch repair mutants[J]. Proceedings of the National Academy of Sciences, 2003, 100(26): 15748-15753.
[11] Hou S, Chen X, Wang H, et al. Efficient method to generate homologous recombinant baculovirus genomes in E. coli[J]. Biotechniques, 2002, 32(4): 783-784, 786, 788.
[12] Hou S, Chen X, Wang H, et al. An efficient method of constructing homologous recom binant baculovirus with PCR amplified fragments[J]. Sci China C: Life Sci, 2003, 46(4): 431-437.
[13] Kitts P A, Ayres M D, Possee R D. Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors[J]. Nucleic Acids Research, 1990, 18(19): 5667-5672.
[14] Kitts P A, Possee R. A method for producing recombinant baculovirus expression vectors at high frequency[J]. Biotechniques, 1993, 14(5): 810-817.

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备注/Memo

备注/Memo:
收稿日期: 2014-01-08
基金项目: 国家高技术研究发展计划“863”项目(2011AA100603)
作者简介: 梅文枫(1988-),女,江西南昌人,硕士研究生,研究方向为生物反应器与蛋白质组学
通信作者: 陈健,电子邮箱:chj1999@126.com
更新日期/Last Update: 2014-11-19