Kit and method for rapidly detecting endocrine disrupting activity of chemical substance based on magnetic microspheres
An endocrine disruptor and chemical substance technology, which is applied in the field of detection of the disrupting effect of chemicals on the human endocrine system, can solve the problem that it is difficult to meet the endocrine disruptor receptor activity of chemicals, high sensitivity and high efficiency are not taken into account, and the experiment repeatability is not good. and other problems, to achieve the effect of short experimental period, strong activity, fast and efficient detection
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[0037] figure 1 It is the whole experimental step process of using the method provided by the invention to detect the endocrine disrupting activity of chemical substances, and the specific experimental operation is described in detail below:
[0038] 1. Preparation and storage of SRC-BAP protein solution
[0039] Using the cDNA of human rectal adenocarcinoma cell Caco-2 as the template, the primers for amplifying the human nuclear receptor transcriptional coactivator SRC were designed, and the SRC fragment was obtained by PCR amplification. The primers of Escherichia coli alkaline phosphatase BAP were increased, and the BAP fragment was obtained by PCR amplification; the SRC fragment and the BAP fragment were jointly ligated to the pET28a vector by double digestion, and the positive clones were screened and sequenced, and the recombinant plasmid was extracted and transformed into the large intestine. Bacillus BL21 to obtain recombinant expression strains. The recombinant str...
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