Objective: To prepare a high purity target protein by expressing Candida albicans acyl carrier protein Acp1 in E. coli BL21. Methods: The target gene Acp1 was amplified by PCR, and then the 6XHis tag and terminator were ligated into the C-terminus of the sequence. After splicing, the sequence was inserted into the plasmid to form the prokaryotic expression vector pET30a-ACP1, which was then transferred into E. coli BL21. IPTG was added to induce the expression. The expression product was collected and purified by the column Ni, and finally the target protein content was measured by the Bradford method. Results: The Candida albicans acyl carrier protein Acp1 was highly expressed in E. coli BL21. The electrophoresis results showed that there were two bands of Acp1 under reducing conditions. After reduction and non-reduction electrophoresis and Western blotting, the results showed that the protein was aggregated in a non-reduced state. The protein Acp1 concentration was determined to be 2.68 mg/mL by the Bradford method. Conclusions: The Candida albicans acyl carrier protein Acp1 is successfully prepared, which provides a basis for the later screening of clinical drugs targeting Ppt2 by fluorescence polarization.
MENG Lingning, LIU Jinyan, WANG Yuting, XIANG Mingjie
. Preparation of recombinant protein acyl carrier protein 1(Acp1) of Candida albicans[J]. Journal of Diagnostics Concepts & Practice, 2018
, 17(06)
: 645
-649
.
DOI: 10.16150/j.1671-2870.2018.06.005
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