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Construction and application of a yeast expression system for thymosin a1
Chen, Feng; Chen, Xiang-Ming; Chen, Zhi; Jiang, Han-Liang; Pan, Xiao-Ping; Hu, Zhong-Rong; Liu, Rong-Hua; Chen, Xiao-Ming.
Afiliação
  • Chen, Feng; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Chen, Xiang-Ming; Medical College of Zhejiang University. 2nd Affiliated Hospital. Department of Anesthesiology. Hangzhou. CN
  • Chen, Zhi; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Jiang, Han-Liang; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Pan, Xiao-Ping; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Hu, Zhong-Rong; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Liu, Rong-Hua; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
  • Chen, Xiao-Ming; Medical College of Zhejiang University. 1st Affiliated Hospital. Institute of Infectious Diseases. Hangzhou. CN
Biocell ; Biocell;29(3): 253-259, Aug.-Dec. 2005. ilus, tab
Article em En | LILACS | ID: lil-633231
Biblioteca responsável: AR1.2
ABSTRACT
We want to construct a yeast expression system for thymosin a1 (Ta1) to make the orally administered Ta1 preparation possible. The whole Ta1 DNA fragment was obtained by PCR. After being digested with restriction enzymes, it was cloned into pYES2 vector. Sequencing was performed to identify the recombinant. The sequence of Ta1 in recombinant coincided with the original one reported in Genbank. When pYES2-Ta1 plasmid was transformed into yeast, galactose instead of glucose was used to induce Ta1 expression. Western blot was performed to identify the quality of the expressed Ta1. Dried yeast containing pYEST2-Ta1 was fed to Balb/c mice whose immunities were inhibited by cyclophosphamide in advance. Synthesized Ta1 peptide was used as positive control and empty yeast was used as negative control. Compared with the negative control group, both dried yeast containing pYEST2-Ta1 and synthesized Ta1 peptide can significantly increase the CD8+ level (22.74±1.09 and 18.77±4.72 vs 7.49±2.14, p<0.01), while both of them had little effect on the CD4+ lymphocytes (61.86±6.94 and 65.91±4.78 vs 57.93±10.40, p>0.05). We concluded that a high effective yeast expression system for Ta1 was constructed successfully and the Ta1 protein expressed by this system can improve CD8+ level in immune inhibited mice.
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Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Saccharomyces cerevisiae / Timosina / Expressão Gênica Limite: Animals Idioma: En Revista: Biocell Assunto da revista: C‚lulas Ano de publicação: 2005 Tipo de documento: Article / Project document País de afiliação: China País de publicação: Argentina
Texto completo: 1 Coleções: 01-internacional Base de dados: LILACS Assunto principal: Saccharomyces cerevisiae / Timosina / Expressão Gênica Limite: Animals Idioma: En Revista: Biocell Assunto da revista: C‚lulas Ano de publicação: 2005 Tipo de documento: Article / Project document País de afiliação: China País de publicação: Argentina