|本期目录/Table of Contents|

[1]徐鹏,张佑红*,杨益,等.流式细胞术快速检测杆状病毒滴度[J].武汉工程大学学报,2010,(01):57-60.
　XU Peng,YANG Yi,PENG Ji ming,et al.Rapid titration of baculovirus by flow cytometry[J].Journal of Wuhan Institute of Technology,2010,(01):57-60.
点击复制

流式细胞术快速检测杆状病毒滴度(/HTML)
分享到：

《武汉工程大学学报》[ISSN:1674-2869/CN:42-1779/TQ]

卷:
期数:
2010年01期
页码:
57-60
栏目:
化学与化学工程
出版日期:
2010-01-31

文章信息/Info

Title:
Rapid titration of baculovirus by flow cytometry
文章编号:
16742869(2010)01005704
作者:
徐鹏; 张佑红*; 杨益; 彭继明; 陈龙; 靖志强; 危威; 马静; 秦琴
武汉工程大学化工与制药学院，湖北 武汉 430074
Author(s):
XU Peng;  YANG Yi;  PENG Jiming;  CHEN Long;  JING Zhiqiang;  WEI Wei;  MA Jing;  QIN Qin;  ZHANG Youhong
School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology, Wuhan 430074, China
关键词:
流式细胞术; 杆状病毒; SYBR Green I
Keywords:
flow cytometry;  baculovirus;  SYBR Green I
分类号:
Q331
DOI:
-
文献标志码:
A
摘要:
为了快速而准确的测定杆状病毒的滴度，采用流式细胞术对经SYBR Green I染色后的杆状病毒直接计数.考察固定、破膜和染色等因素对检测结果的影响，并验证改进后的方法的重复性和线性性.杆状病毒最佳染色条件：质量分数为0.1%的多聚甲醛固定病毒30 min后，加入一定量的SYBR Green I在80 ℃下孵育10 min.测量结果CV值为2.4%(n=8)，R2为0.999 8.整个测量过程由原来终点稀释法的7~10 d缩短到1 h.
Abstract:
To develop a rapid and accurate method for baculovirus titration, flow cytometry (FCM) was adopted to quantitate baculovirus directly after staining with SYBR Green I. The effects of different treatment steps (fixation, permeabilization, staining, etc.) were examined. The linearity and the reproducibility of the developed method were evaluated. The optimum dyeing conditions were shown: viral samples fixed with 0.1% paraformaldehyde for 30 min at 4 ℃, stained with SYBR Green I (1×10-4 commercial concentration) for 10 min at 80 ℃. The CV value of results is 2.4%(n=8), R2 value of different dilutions is 0.999 8. The entire procedure is shortened to 1 h compared with the 7-10 d of end-point dilution method.

参考文献/References:

1］Thomas A K, Condreay J P, Donald L J. Baculovirus as versatile vectors for protein expression in insect and mammalian cells［J］. Nat Biotechnol, 2005, 23(5): 567575.
［2］Radford K M, Cavegn C, Bertrand M, et al. The indirect effects of multiplicity of infection on baculovirus expressed proteins in insect cells secreted and nonsecreted products［J］. Cytotechnology, 1997, 24: 7381.
［3］Zhang Y H, Enden G, Merchuk J C. Insect cellsBaculovirus system: Factors affecting growth and low MOI infection［J］. Biochem Eng J, 2005, 27(1): 816.
［4］O’Reilly D R, Miller L K, Luckow V A. Baculovirus Expression Vectors: A Laboratory Manual［M］. New York: Oxford University Press, 1994: 132134.
［5］Hink W F, Vail P V. A plaque assay for titration of Alfalfa looper nuclear polyhedrosis virus in cabbage looper (TN-368) cell line［J］. J Invertebr Pathol, 1973, 22: 168174.
［6］Marie D, Brussaard C P D, Thyrhaug R, et al. Enumeration of marine viruses in culture and natural samples by flow cytometry［J］. Appl Environ Microbiol, 1999, 65(1): 4552.
［7］Brussaard C P D. Optimization of procedures for counting viruses by flow cytometry［J］. Appl Environ Microbiol, 2004, 70(3): 15061513.
［8］Shen C F, Meghrous J, Kamen A. Quantitation of baculovirus particles by flow cytometry［J］. J Virol Methods, 2002, 105(2): 321330.
［9］Fineey D J. Statistical Method in Biological Assay［M］. 3rd ed. London: Charles Griffin & Co., 1978: 394401

相似文献/References:

[1]张佑红,杨文,危威,等.杆状病毒在气升式反应器中感染策略优化[J].武汉工程大学学报,2012,(06):22.[doi:103969/jissn16742869201206006]
　ZHANG You\|hong,YANG Wen,WEI Wei,et al.Optimization of infection strategy for baculovirus in airlift reactor[J].Journal of Wuhan Institute of Technology,2012,(01):22.[doi:103969/jissn16742869201206006]

备注/Memo

备注/Memo:
收稿日期：20091022基金项目：国家自然科学基金(20876120)资助项目作者简介：徐鹏(1985)，男，四川内江人，硕士研究生.研究方向： 细胞生物学.指导老师：张佑红，男，博士，教授，硕士研究生导师.研究方向：细胞生物学、分子生物学.*通信联系人
更新日期/Last Update: