Construction and application of a yeast expression system for thymosin alpha1
| Autor: | Feng, Chen, Xiang-Ming, Chen, Zhi, Chen, Han-Liang, Jiang, Xiao-Ping, Pan, Zhong-Rong, Hu, Rong-Hua, Liu, Xiao-Ming, Chen |
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| Rok vydání: | 2006 |
| Předmět: |
Mice
Inbred BALB C Thymalfasin T-Lymphocytes Blotting Western Genetic Vectors CD4-CD8 Ratio Gene Expression Saccharomyces cerevisiae CD8-Positive T-Lymphocytes Flow Cytometry Polymerase Chain Reaction Recombinant Proteins Clone Cells Thymosin Mice Random Allocation Sonication Freeze Drying Animals Cloning Molecular Cyclophosphamide Immunosuppressive Agents Injections Intraperitoneal |
| Zdroj: | Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al. 29(3) |
| ISSN: | 0327-9545 |
| Popis: | We want to construct a yeast expression system for thymosin alpha1 (Talpha1) to make the orally administered Talpha1 preparation possible. The whole Talpha1 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 Talpha1 in recombinant coincided with the original one reported in Genbank. When pYES2-Talpha1 plasmid was transformed into yeast, galactose instead of glucose was used to induce Talpha1 expression. Western blot was performed to identify the quality of the expressed Talpha1. Dried yeast containing pYEST2-Talpha1 was fed to Balb/c mice whose immunities were inhibited by cyclophosphamide in advance. Synthesized Talpha1 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-Talpha1 and synthesized Talpha1 peptide can significantly increase the CD8+ level (22.74 +/- 1.09 and 18.77 +/- 4.72 vs 7.49 +/- 2.14, p0.01), 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,p0.05). We concluded that a high effective yeast expression system for Talpha1 was constructed successfully and the Talpha1 protein expressed by this system can improve CD8+ level in immune inhibited mice. |
| Databáze: | OpenAIRE |
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