• Title/Summary/Keyword: laboratory

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New Hydroperoxides from Aster oharai

  • Park, Sang-Zin;Kwon, Hak-Cheol;Min, Yong-Deuk;Lee, Sung-Ock;Lee, Won-Bin;Yang, Min-Cheol;Chung, Ae-Kyung;Lee, Kang-Ro
    • Proceedings of the PSK Conference
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    • 2001.10a
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    • pp.258.2-259
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    • 2001

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Codon Optimization, Soluble Expression and Purification of PE_PGRS45 Gene from Mycobacterium tuberculosis and Preparation of Its Polyclonal Antibody Protein

  • Xu, Tao;Li, Minying;Wang, Chutong;Yuan, Meili;Chang, Xianyou;Qian, Zhongqing;Li, Baiqing;Sun, Meiqun;Wang, Hongtao
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1583-1590
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    • 2021

  • Studies have demonstrated that PE_PGRS45 is constitutively expressed under various environmental conditions (such as nutrient depletion, hypoxia, and low pH) of the in vitro growth conditions examined, indicating that PE_PGRS45 protein is critical to the basic functions of Mycobacterium tuberculosis. However, there are few reports about the biochemical function and pathogenic mechanism of PE_PGRS45 protein. The fact that this M. tuberculosis gene is not easily expressed in E. coli may be mainly due to the high content of G+C and the use of unique codons. Fusion tags are indispensable tools used to improve the soluble expression of recombinant proteins and accelerate the characterization of protein structure and function. In the present study, His6, Trx, and His6-MBP were used as fusion tags, but only MBP-PE_PGRS45 was expressed solubly. The purification using His6-MBP tag-specific binding to the Ni column was easy to separate after the tag cleavage. We used the purified PE_PGRS45 to immunize New Zealand rabbits and obtained anti-PE_PGRS45 serum. We found that the titer of polyclonal antibodies against PE_PGR45 was higher than 1:256000. The result shows that purified PE_PGRS45 can induce New Zealand rabbits to produce high-titer antibodies. In conclusion, the recombinant protein PE_PGRS45 was successfully expressed in E. coli and specific antiserum was prepared, which will be followed by further evaluation of these specific antigens to develop highly sensitive and specific diagnostic tests for tuberculosis.

  • https://doi.org/10.4014/jmb.2106.06006 인용 PDF KSCI