Background[1-3]
Aminoglycoside phosphate domain protein antibodies are a type of polyclonal antibody that can specifically bind to aminoglycoside phosphate domain proteins. They are mainly used for Western Blot, IHC-P, and IF detection of aminoglycoside phosphate domain proteins. , ELISA, Co-IP and other immunological experiments.
Detection principle: Double-antibody sandwich method determines the level of aminoglycoside phosphate domain protein in the specimen. Use the purified aminoglycoside phosphate domain protein antibody to coat the microwell plate to make a solid-phase antibody. Add the aminoglycoside phosphate domain protein to the microwells coated with the monoclonal antibody in sequence, and then mix it with HRP-labeled aminoglycoside phosphate. The domain protein antibody binds to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme and into the final yellow under the action of acid. The depth of the color is positively correlated with the aminoglycoside phosphate domain protein in the sample. Use a microplate reader to measure the absorbance (OD value) at a wavelength of 450 nm, and calculate the concentration of aminoglycoside phosphate domain protein in the sample through the standard curve.
Aminoglycosides are glycoside antibiotics composed of amino sugars and amino cyclic alcohols connected through an oxygen bridge. Aminoglycoside antibiotics inhibit protein synthesis and are stationary phase bactericidal antibiotics. It is characterized by its resistance to aerobic Gram-negative bacilli, Pseudomonas, Mycobacterium and Staphylococcus aureus. Since aminoglycoside antibiotics must have oxygen when they exert their antibacterial effects, they are ineffective against anaerobic bacteria.
Aminoglycoside antibiotics are glycosides formed from amino sugars (monosaccharides or double sugars) and amino alcohols. Because they contain amino groups and other basic groups, these antibiotics are alkaline and are usually prepared into crystalline sulfate or hydrochloride in clinical practice. This type of antibiotic contains multiple hydroxyl groups and is a polar compound with high water solubility and low fat solubility. When administered orally, the absorption is less than 10% and must be administered by injection. Its aqueous solution is very stable in the pH range of 2 to 11. Since there are several chiral carbons in the sugar part of the molecule, it is optically active, and the binding rate of these antibiotics to serum proteins is low. Most of them are not metabolized and inactivated in the body, and are excreted through glomerular filtration in the form of the original drug. Toxic to the kidneys.
Apply[4][5]
Pharmacodynamic study of the new aminoglycoside antibiotic vertilmicin and its stability study on the recombinant aminoglycoside bifunctional modification enzyme
The main way that bacteria are resistant to aminoglycoside antibiotics is to produce aminoglycoside-modifying enzymes, which modify the groups necessary to maintain antibacterial activity in the drug molecules, greatly reducing their affinity with the target ribosome. . The most common modifying enzyme in Gram-positive bacteria is the aminoglycoside bifunctional modifying enzyme AAC (6′)-APH (2″). This enzyme has both N-terminal acetyltransferase activity and C-terminal phosphotransferase activity. , can modify almost all types of aminoglycoside antibiotics to eliminate their synergistic effect with antibiotics that act on cell walls, such as penicillin.
By recombinantly expressing aminoglycoside bifunctional modification enzymes AAC (6′), APH (2″) and AAC (6′)-APH (2″) in Escherichia coli, preparing enzyme extracts and constructing recombinant aminoglycosides In vitro metabolism model of bifunctional modifying enzymes, the stability of acetyltransferase activity and phosphotransferase activity of vertilmicin on aminoglycoside bifunctional modifying enzymes was studied, and by comparing AAC(6′)-APH(2 ″), AAC (6′), and APH (2″) on the modification effects of antibiotics Japan Tosoh Matting Agent Price were preliminarily studied on the effect of gene fusion on enzyme modification activity.
