Establishment of an avian leukosis virus subgroup A-resistant cell line

doi:10.1016/S2095-3119(16)61453-3

Journal of Integrative Agriculture ›› 2017, Vol. 16 ›› Issue (04): 930-936.DOI: 10.1016/S2095-3119(16)61453-3

收稿日期:2016-04-13 出版日期:2017-04-04 发布日期:2017-04-07

Establishment of an avian leukosis virus subgroup A-resistant cell line

FENG Min^{1, 2, 3, 4*}, DAI Man-man^{1, 2, 3, 4*}, LIAO Ming^{1, 2, 3, 4}, CAO Wei-sheng^{1, 2, 3, 4}

¹College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China
²National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China
³Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China
⁴Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.China

Received:2016-04-13 Online:2017-04-04 Published:2017-04-07
Contact: CAO Wei-sheng, Tel: +86-20-85282536, Fax: +86-20-85280234, E-mail: caoweish@scau.edu.cn
About author:FENG Min, E-mail: hunanfengmin@163.com
Supported by:
The work was founded by the National Key R&D Program of China (2016YFD0501606), the Public Industry Research Program, the Ministry of Agriculture of China (201203055), the Program of Science and Technology Development of Guangdong Province, China (2015A020209145), the China Meat-Type Chicken Research System (CARS-42-G09) and the Modern Agriculture Talents Support Program, Ministry of Agriculture of China ([2012] no.160).

摘要/Abstract

Abstract: Rapid diagnostic methods for classifying avian leukosis subgroups in the field were needed for routine, large-scale screening. As a first step in method development, we inserted the avian leukosis virus subgroup A (ALV-A) env gene into plasmid pcDNA3.1/Zeo (+) and used this construct to transfect DF-1 cells. Zeocin-resistant cells were obtained after 2 weeks of zeocin selection. Then, the cells were analyzed using PCR, immunofluorescence, and Western blot for expression of the envA-encoded envelope protein after 30 serial passages. The DF-1/A cell line was completely resistant to 10⁴TCID₅₀/0.1 mL (50% tissue culture infective dose)ALV-A and was partially resistant to 10⁵ TCID₅₀/0.1 mL ALV-A viral particles. By comparing the DF-1/A and DF-1 cell lines, an ALV-A isolate was identified using a gag-specific ELISA for capsid protein p27. Thus, we established a DF-1/A cell line that was resistant to ALV-A infection. This cell line will be useful as a diagnostic tool.

Key words: avian leukosis virus subgroup A (ALV-A), env, DF-1/A cell line, diagnostic tool

. [J]. Journal of Integrative Agriculture, 2017, 16(04): 930-936.

FENG Min, DAI Man-man, LIAO Ming, CAO Wei-sheng. Establishment of an avian leukosis virus subgroup A-resistant cell line[J]. Journal of Integrative Agriculture, 2017, 16(04): 930-936.

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Establishment of an avian leukosis virus subgroup A-resistant cell line

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