Genetic engineering kit for detecting environmental estrogen pollutants and application thereof
A genetic engineering and estrogen technology, which is applied in the field of genetic engineering kits for the detection of environmental estrogen pollutants, can solve the problems of high false-positive antigen-antibody reaction, high price and high cost, and achieves a simple and fast detection process and reduces analysis time. The effect of low cost and sample requirements
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0033] Preparation of genetic engineering kit
[0034] Step 1, fusing the Ser301-Thr553 peptide segment of human estrogen receptor alpha protein with Mycobacterium tuberculosis recA intein to construct an estrogen-sensitive intein chimera;
[0035] (1) PCR amplified intein (Mycobacterium tuberculosis recA intein) fragment, primers were 5'-GGATCCtgtttcgactacagcacccg-3', 5'-AAGCTTgttgtgcaccatgacaccgt-3', purified and recovered; the fragment and plasmid pET28a were respectively used with Bam HI / After Hind III double digestion, the restriction fragment was recovered; then the two restriction fragments were ligated with T4 DNA ligase, and the Escherichia coli DH5α (F-endA1gln V44thi-1recA1relA1gyrA96deoRnupGΦ80dlacZΔM15Δ(lacZYA-argF)U169, hsdR17(rK-mK+), hsdR17(rK-mK+), λ-), spread LBK plates, culture overnight at 37°C; pick clones and extract plasmids, sequence and identify, positive plasmids were named pMRecA, purified and recovered, and stored at -20°C for future use.
[0036]...
Embodiment 2
[0061] Detection of Effect Levels of Various Environmental Estrogens by Genetic Engineering Kit
[0062] Using the kit obtained in Example 1, different environmental estrogen samples and sewage samples obtained from sewage treatment plants were measured, and the estrogen effect level of the samples was analyzed.
[0063] Prepare different concentrations of 17β-estradiol, bisphenol A, chrysene and 6-OH- -1 , 1×10 -2 , 1×10 -3 , 1×10 -4 , 1×10 -5 , 1×10 -6 , 1×10 -7 , 1×10 -8 , 1×10 -9 , 1×10 -10 mg / mL) samples, stored at low temperature. The sewage sludge sample obtained from the sewage treatment plant was filtered with filter paper, and the sample to be tested was obtained by solid phase extraction. When measuring samples, inoculate estrogen-sensitive genetically engineered strains into 1mL liquid medium, shake at 37°C until OD 600 At about 0.6, add 1 μL of inducer (1:1000 dilution) and 10 μL of sample solutions with different concentrations. Shake culture at 37°C ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com