An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced a...An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced antibody reactions specific to subgroup A or B. But gp85 amino acid sequence comparisons indicated that SDAU09C1 fell into subgroup A; it had homology of 88.8%-90.3% to 6 reference strains of subgroup A, much higher compared to other subgroups including subgroup B. This is the first report for ALV of subgroup A isolated from imported breeders.展开更多
Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without a...Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without anti-ALV-J serum in the medium); B (with anti-ALV-J serum in the medium), then viruses from each well of both groups were separately passed in CEF every 6 d; formed their independent passage lineages. For each lineage of both groups, gp85 genes of the viruses in the 10th, 20th; 30th passages were amplified, cloned; sequenced. The sequence data indicated that the homologies of gp85 at aa level between the primary virus; the passed viruses of different passages of 3 lineages in group A were 97.7%–99.7%;; the homologies of gp85 between the primary virus; the passed viruses of different passages of 3 lineages in group B were 93.8%–96.1%. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S in 3 highly variable (hr-) regions at aa#110–120, aa#141–151; aa#189–194 of gp85 in 3 lineages of group A were 2 (8/4), 1(3/3); 1.3 (4/3), however, NS/S in the same 3 hr-regions of group B were 4.1 (13/3), 4.7 (14/3); 3.3 (11/3). This study is the first demonstration of influence of immune selective pressure on evolution of ALV-J gp85 by specific antibodies under the controlled in vitro experiments.展开更多
Blood samples were collected from a local strain of chickens associated with serious tumor cases in Shandong Province. The samples were inoculated into chicken embryo fibroblast and DF-1 cells for virus isolation and ...Blood samples were collected from a local strain of chickens associated with serious tumor cases in Shandong Province. The samples were inoculated into chicken embryo fibroblast and DF-1 cells for virus isolation and identification, respectively. The inoculated cells were screened for three common chicken tumor viruses. Nine strains of avian leukosis virus subgroup J （ALV-J） were identified, and were designated LY1201-LYI209. The env gene from the LY1201 strain was amplified and cloned. All nine resultant env clones （clones 01-09） were sequenced, and the gp85 and gp37 amino acid regions were subjected to homology analysis. Clones 01 and 03 had 10 amino acid deletions in the gp85 region compared to the other seven clones, suggesting that at least two quasispecies with obvious mutations coexist in the same field strain. Among these nine clones, three had identical gp85 and gp37 sequences, and were recognized as the dominant LY1201 quasispecies. The amino acid sequence homology of gp37 and gp85 among the nine clones was 98.5%-100.0% and 96.6%-100.0% respectively, suggesting that the gp85 region of the env gene can better display the quasispecies diversity of ALV-J than gp37.展开更多
[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identifi...[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identified by pathological anatomy, DF- 1 cell culture and RT- PCR. And then they were named HB1002, HB1003 and HB1009, respectively. [ Result] Test sequence analysis showed, the length of gp85 gene was 921 bp, consistent with expect result; the nucleotide homology between the three isolates was in 97.7% -99.7% ,the homology of amino acid was in 95.1% - 99%. The nucleotide homology between HPRS - 103 and the three isolates was in the 94.1% - 94.8% ; and the nucleotide homolo- gy between other ALV -J and the three isolates was in 87.6% -97.3%. The phylogenetic trees analysis showed that the homology of JS09GY6 vi- rus and the three isolates was nearest, in 95.2% -97.3%. [ Conclusion] In the test, the three strains of virus which were isolated were ALV- J.展开更多
The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This repor...The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).展开更多
基金supported by a grant from Shandong province and a grant (200803019) from the Ministry of Agriculturesupported by a grant from Shandong province and a grant (20080019) from the Ministry of Agriculture
文摘An exogenous avian leukosis virus (ALV) strain SDAU09C1 was isolated in DF-1 cells from one of 240 imported 1-day-old white meat-type grand parent breeder chicks. Inoculation of SDAU09C1 in ALV-free chickens induced antibody reactions specific to subgroup A or B. But gp85 amino acid sequence comparisons indicated that SDAU09C1 fell into subgroup A; it had homology of 88.8%-90.3% to 6 reference strains of subgroup A, much higher compared to other subgroups including subgroup B. This is the first report for ALV of subgroup A isolated from imported breeders.
文摘Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without anti-ALV-J serum in the medium); B (with anti-ALV-J serum in the medium), then viruses from each well of both groups were separately passed in CEF every 6 d; formed their independent passage lineages. For each lineage of both groups, gp85 genes of the viruses in the 10th, 20th; 30th passages were amplified, cloned; sequenced. The sequence data indicated that the homologies of gp85 at aa level between the primary virus; the passed viruses of different passages of 3 lineages in group A were 97.7%–99.7%;; the homologies of gp85 between the primary virus; the passed viruses of different passages of 3 lineages in group B were 93.8%–96.1%. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S in 3 highly variable (hr-) regions at aa#110–120, aa#141–151; aa#189–194 of gp85 in 3 lineages of group A were 2 (8/4), 1(3/3); 1.3 (4/3), however, NS/S in the same 3 hr-regions of group B were 4.1 (13/3), 4.7 (14/3); 3.3 (11/3). This study is the first demonstration of influence of immune selective pressure on evolution of ALV-J gp85 by specific antibodies under the controlled in vitro experiments.
基金financially supported by a special grant from Ministry of Agriculture of China in 2012
文摘Blood samples were collected from a local strain of chickens associated with serious tumor cases in Shandong Province. The samples were inoculated into chicken embryo fibroblast and DF-1 cells for virus isolation and identification, respectively. The inoculated cells were screened for three common chicken tumor viruses. Nine strains of avian leukosis virus subgroup J （ALV-J） were identified, and were designated LY1201-LYI209. The env gene from the LY1201 strain was amplified and cloned. All nine resultant env clones （clones 01-09） were sequenced, and the gp85 and gp37 amino acid regions were subjected to homology analysis. Clones 01 and 03 had 10 amino acid deletions in the gp85 region compared to the other seven clones, suggesting that at least two quasispecies with obvious mutations coexist in the same field strain. Among these nine clones, three had identical gp85 and gp37 sequences, and were recognized as the dominant LY1201 quasispecies. The amino acid sequence homology of gp37 and gp85 among the nine clones was 98.5%-100.0% and 96.6%-100.0% respectively, suggesting that the gp85 region of the env gene can better display the quasispecies diversity of ALV-J than gp37.
基金Plan of Hubei Province Science and Technology Research and Development Project:the Research and Demonstration of Comprehensive Prevention and Control Technology on Poultry Avian Influenza and other Major Diseases YJN0065the Construction Fund of Modern Agriculture Industry Technology System CARS-42-G11
文摘[Objective] To isolate three strains of avian leukosis virus subgroup J（ALV -J） , and then amplify and sequence the gp85 gene. [ Method] Three strains of ALV- J were isolated from Hubei Province, which were identified by pathological anatomy, DF- 1 cell culture and RT- PCR. And then they were named HB1002, HB1003 and HB1009, respectively. [ Result] Test sequence analysis showed, the length of gp85 gene was 921 bp, consistent with expect result; the nucleotide homology between the three isolates was in 97.7% -99.7% ,the homology of amino acid was in 95.1% - 99%. The nucleotide homology between HPRS - 103 and the three isolates was in the 94.1% - 94.8% ; and the nucleotide homolo- gy between other ALV -J and the three isolates was in 87.6% -97.3%. The phylogenetic trees analysis showed that the homology of JS09GY6 vi- rus and the three isolates was nearest, in 95.2% -97.3%. [ Conclusion] In the test, the three strains of virus which were isolated were ALV- J.
基金Natural Science Foundation of China (30460098)China Postdoctoral Science Foundation funded project (2005038585)
文摘The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).
