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Journal of Integrative Agriculture  2017, Vol. 16 Issue (04): 930-936    DOI: 10.1016/S2095-3119(16)61453-3
Animal Science · Veterinary Science Advanced Online Publication | Current Issue | Archive | Adv Search |
Establishment of an avian leukosis virus subgroup A-resistant cell line
FENG Min1, 2, 3, 4*, DAI Man-man1, 2, 3, 4*, LIAO Ming1, 2, 3, 4, CAO Wei-sheng1, 2, 3, 4

1 College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China

2 National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China

3 Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China

4 Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.China

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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 104 TCID50/0.1 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.
Keywords:  avian leukosis virus subgroup A (ALV-A)      env      DF-1/A cell line      diagnostic tool  
Received: 13 April 2016   Accepted:

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).

Corresponding Authors:  CAO Wei-sheng, Tel: +86-20-85282536, Fax: +86-20-85280234, E-mail:   
About author:  FENG Min, E-mail:

Cite this article: 

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

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