Construction of a eukaryotic expression system with stable and secretory expression of mycobacterium tuberculosis 38 kDa protein.

Autor: Chen H; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China.; Sun Yat-Sen Memorial Hospital, Sun Yat-Sen Univsity, Guangzhou, 510000, China., Chen Z; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China., Bai N; Department of Nuclear Medicine, Yuxi People's Hospital of Yunnan Province, Yuxi, 653100, China., Yan R; Guangzhou Bioneeds Biotechnology CO., LTD, Guangzhou, 510000, China., Xu M; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China., Wu W; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China.; Animal Science and Technology College, Jilin Agricultural University, Changchun, 130118, China., Liang W; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China., Li H; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China. hongwei1@yahoo.com., Mao Y; School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China. maoyy2011@163.com.
Jazyk: angličtina
Zdroj: World journal of microbiology & biotechnology [World J Microbiol Biotechnol] 2021 Sep 14; Vol. 37 (10), pp. 175. Date of Electronic Publication: 2021 Sep 14.
DOI: 10.1007/s11274-021-03143-x
Abstrakt: The 38 kDa protein is a major antigen of mycobacterium tuberculosis and has been widely used in TB serodiagnosis, due to its highly sensitivity and specificity. Here we attempt to establish a production platform of recombinant 38 kDa protein in mammalian cells and to evaluate the potential value of 38 kDa protein in TB serodiagnosis. The 38 kDa gene is synthesized and cloned into a lentiviral expressing vector. Recombinant lentiviral vector LV-CMV-38 kDa-eGFP was packaged, titered, and then transduced into HEK 293 T cells. Recombinant cell lines were selected by limiting dilution. Supernatants were collected and purified by HisTrapTM HP column. Western blot showed a molecular weight of approximate 38 kDa in cell supernatants as expected. ELISA assay confirmed the immunological specificity of the obtained protein in the presence of MTB-infected human serum samples. In all, we have obtained a stable cell line with long-term and robust expression of secretory MTB 38 kDa protein, which may provide a promising candidate antigen for the development of TB serological diagnosis.
(© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)
Databáze: MEDLINE
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