Cloning, Expression And Purification Of Acidic Mammalian Chitinase And Its Hydrolysis Of Some Fungal Cell Wall

Opportunistic fungal infection in immunocompromised patients is a major and important problem in our clinical practice. Antifungal agents are challenged by their hepatorenal toxicity, drug resistance and single antimicrobial spectrum. So it is urgent to develop effective, safe and broad-spectrum antifungal agents.Chitin, being present as a structural component in cell wall of fungus, plays an important role in the metabolism. There is no chitin in human, so acidic mammalian chitinase (AMCase) is a safe and promising antifungal agent.OBJECTIVE:To clone AMCase gene and analyze its antifungal characteristics with bioinformation technology. To express AMCase protein and extract it from the stomach of BALB/c mouse. To identify its suppressive function to Candida albicans, Aspergillus fumigatus and Trichophyton rubrum with agar diffusion testing and In vitro susceptibility testing, et al. giving insights into the development of non-noxious antifungal agents.METHODS:1. Total RNA was extracted from the stomach of BALB/c mouse using Trizol reagent and was reversely transcribed into cDNA. AMCase gene was...