Establishment of an avian leukosis virus subgroup A-resistant cell line
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...
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KeAi Communications Co., Ltd.
2017-04-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311916614533 |
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| author | Min FENG Man-man DAI Ming LIAO Wei-sheng CAO |
| author_facet | Min FENG Man-man DAI Ming LIAO Wei-sheng CAO |
| author_sort | Min FENG |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e95d97fdc0394107b2ff598a21b4aa2e |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2017-04-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-e95d97fdc0394107b2ff598a21b4aa2e2025-08-20T03:57:52ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192017-04-0116493093610.1016/S2095-3119(16)61453-3Establishment of an avian leukosis virus subgroup A-resistant cell lineMin FENG0Man-man DAI1Ming LIAO2Wei-sheng CAO3College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China; Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China; Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China; Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.ChinaCollege of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, P.R.China; National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Guangzhou 510642, P.R.China; Key Laboratory of Veterinary Vaccine Innovation, Ministry of Agriculture, Guangzhou 510642, P.R.China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou 510642, P.R.China; Correspondence CAO Wei-sheng, Tel: +86-20-85282536, Fax: +86-20-85280234Rapid 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.http://www.sciencedirect.com/science/article/pii/S2095311916614533avian leukosis virus subgroup A (ALV-A)envDF-1/A cell linediagnostic tool |
| spellingShingle | Min FENG Man-man DAI Ming LIAO Wei-sheng CAO Establishment of an avian leukosis virus subgroup A-resistant cell line Journal of Integrative Agriculture avian leukosis virus subgroup A (ALV-A) env DF-1/A cell line diagnostic tool |
| title | Establishment of an avian leukosis virus subgroup A-resistant cell line |
| title_full | Establishment of an avian leukosis virus subgroup A-resistant cell line |
| title_fullStr | Establishment of an avian leukosis virus subgroup A-resistant cell line |
| title_full_unstemmed | Establishment of an avian leukosis virus subgroup A-resistant cell line |
| title_short | Establishment of an avian leukosis virus subgroup A-resistant cell line |
| title_sort | establishment of an avian leukosis virus subgroup a resistant cell line |
| topic | avian leukosis virus subgroup A (ALV-A) env DF-1/A cell line diagnostic tool |
| url | http://www.sciencedirect.com/science/article/pii/S2095311916614533 |
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