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Expression and purification of swine RAG2 in E. coli for production of porcine RAG2 polyclonal antibodies.
Jin, Yu-Bei; Yang, Wen-Tao; Huang, Ke-Yan; Chen, Hong-Liang; Shonyela, Seria-Masole; Liu, Jing; Liu, Qiong; Feng, Bo; Zhou, You; Zhi, Shu-Li; Jiang, Yan-Long; Wang, Jian-Zhong; Huang, Hai-Bin; Shi, Chun-Wei; Yang, Gui-Lian; Wang, Chun-Feng.
Afiliação
  • Jin YB; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Yang WT; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Huang KY; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Chen HL; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Shonyela SM; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Liu J; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Liu Q; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Feng B; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Zhou Y; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Zhi SL; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Jiang YL; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Wang JZ; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Huang HB; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Shi CW; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Yang GL; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
  • Wang CF; a College of Animal Science and Technology, Jilin Provincial Engineering Research Center of Animal Probiotics, Key Laboratory of Animal Production and Product Quality Safety of Ministry of Education , Jilin Agricultural University , Changchun , China.
Biosci Biotechnol Biochem ; 81(8): 1489-1496, 2017 Aug.
Article em En | MEDLINE | ID: mdl-28644752
ABSTRACT
Recombination activating gene 2 (RAG2) is necessary for immature B cell differentiation. Antibodies to human and rabbit RAG2 are currently commercially available, but antibodies to swine RAG remain unavailable to date. In this study, the swine RAG2 genes sequence was synthesized and then cloned into a pET-28a vector. The recombinant fusion protein was successfully expressed in E. coli, purified through nickel column chromatography, and further digested with Tobacco Etch Virus protease. The cleaved protein was purified by molecular-exclusion chromatography and named pRAG2. We used pRAG2 to immunize rabbits, collected the serum and purified rabbit anti-pRAG2 polyclonal antibodies. The rabbit anti-pRAG2 polyclonal antibodies were tested via immunofluorescence on eukaryotic cells overexpressing pRAG2 and also able to recognize pig natural RAG2 and human RAG2 protein in western blotting. These results indicated that the prepared rabbit anti-pRAG2 polyclonal antibodies may serve as a tool to detect immature B cell differentiation of swine.
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Texto completo: 1 Coleções: 01-internacional Contexto em Saúde: 3_ND Base de dados: MEDLINE Assunto principal: Proteínas Nucleares / Expressão Gênica / VDJ Recombinases / Proteínas de Ligação a DNA / Escherichia coli / Anticorpos Limite: Animals / Humans Idioma: En Revista: Biosci Biotechnol Biochem Ano de publicação: 2017 Tipo de documento: Article
Texto completo
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Texto completo: 1 Coleções: 01-internacional Contexto em Saúde: 3_ND Base de dados: MEDLINE Assunto principal: Proteínas Nucleares / Expressão Gênica / VDJ Recombinases / Proteínas de Ligação a DNA / Escherichia coli / Anticorpos Limite: Animals / Humans Idioma: En Revista: Biosci Biotechnol Biochem Ano de publicação: 2017 Tipo de documento: Article