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The invention discloses an E3 ubiquitin enzyme mutant as well as modification and a protein targeted degradation system and application thereof, and particularly relates to a series of targeted protein degradation enhanced E3 ubiquitin enzymes (Trim21-N) and a modification method thereof, the E3 ubiquitin enzymes (Trim21-N) are obtained by modifying a wild type Trim21 (wtTrim21) protein structural domain, and the E3 ubiquitin enzymes comprise Trim21-a, Trim21-b, Trim21-c, Trim21-d, Trim21-e and Trim21-f. The efficiency of degrading the target protein through the ubiquitin proteasome is remarkably improved, and the method has the characteristics of strong endogenous and exogenous universality, high specificity, ubiquitin proteasome dependence, low cytotoxicity, strong cell compatibility and the like on the degradation of the target protein. The invention provides E3 ubiquitin enzyme (Trim21-N) based on a targeted protein degradation enhancement version and an exogenous fluorescence report degradation system of the E3 ubiquitin enzyme (Trim21-N), preferably Trim21-e with the optimal degradation effect, and the E3 ubiquitin enzyme (Trim21-N) is named as delta Trim21. The invention further provides an endogenous protein degradation system based on the delta Trim21. The method has the characteristics of high specificity, rapidness and small module, so that the method can be used for researching various biological and disease mechanisms, including the fields of cell targeted therapy, cell mechanism exploration, genetic engineering and the like.

The invention discloses a monoclonal antibody composition for resisting neurofilament light chain protein and application, the monoclonal antibody composition comprises NfL-Clone 1 and NfL-Clone 2, the amino acid sequence of a heavy chain variable region of the NfL-Clone 1 is as shown in SEQ ID NO.1, and the amino acid sequence of a light chain variable region of the NfL-Clone 1 is as shown in SEQ ID NO.2; the amino acid sequence of a heavy chain variable region of the NfL-Clone 2 is as shown in SEQ ID NO.3, and the amino acid sequence of a light chain variable region of the NfL-Clone 2 is as shown in SEQ ID NO.4. The antibody combination has the characteristics of specificity and high affinity, the NfL can be used as a brain injury biomarker, and meanwhile, the invention also provides a diagnostic detection kit of the antibody for early diagnosis of brain injury of a patient, so that the specificity and sensitivity of a diagnostic reagent are improved.

This invention relates to the field of biomedical detection technology, providing a reagent, kit, and detection method for detecting *Vibrio cholerae* serogroup O139 using loop-mediated isothermal amplification (LAMP). This invention targets the synergistic regulation of the tcpA and hlyA genes in *Vibrio cholerae* serogroup O139, designing LAMP detection primers and establishing a LAMP detection method based on isothermal amplification. These LAMP detection primers or loop primers show promising applications not only against *Vibrio cholerae* serogroup O139 but also against other nosocomial infection bacteria. The LAMP primers for detecting *Vibrio cholerae* serogroup O139 can be used to detect *Vibrio cholerae* serogroup O139 in clinical patients or for outpatient screening. The kit of this invention can effectively and specifically detect single-reaction, single-copy, or higher copies of *Vibrio cholerae* serogroup O139. Compared to the "gold standard" real-time quantitative PCR, the kit of this invention offers higher accuracy and sensitivity, completing the detection within 30 minutes.

The invention relates to the field of molecular biology, in particular to a primer group, an MYH3 gene promoter region DNA methylation level analysis method and application. The primer group comprises a first primer pair and a second primer pair, the upstream sequence of the first primer pair is as shown in SEQ ID NO. 1; the downstream sequence of the first primer pair is as shown in SEQ ID NO. 2; the upstream sequence of the second primer pair is as shown in SEQ ID NO.3; and the downstream sequence of the second primer pair is as shown in SEQ ID NO.4. The method for analyzing the methylation level of the DNA in the MYH3 gene promoter region relates to PCR reaction conditions for obtaining qualified target DNA fragments for MSP sequencing, the primer group and the PCR reaction conditions are combined to have the advantage of high amplification efficiency, the qualified DNA fragments are obtained through amplification, the DNA fragments are provided for analyzing the methylation level of the DNA in the MYH3 gene promoter region, and the methylation level analysis efficiency and accuracy are improved.

The application discloses a preparation method and use method of an electrochemical molecular imprinting sensor for detecting DBP. A PEDOT-PG nanocomposite is prepared through a liquid-liquid interface polymerization method, then the prepared PEDOT-PG is functionalized by using polylysine, so that the PEDOT / PG has good dispersity and stability, and good adhesion on the surface of the sensor. In the presence of DBP, a molecular imprinting film is prepared through a monomer electropolymerization method, so that the recognition response speed of the target molecule is improved, the molecular imprinting sensor has the advantages of structural stability, high mechanical strength, easy acquisition and low cost, and has the advantages of specificity for recognizing the target molecule.