Method for high-throughput screening of phage host spectrums

Abstract

The invention discloses a method for high-throughput screening of phage host spectrums and belongs to the technical field of phage host spectrum screening. According to the method disclosed by the invention, the purified bacteriophages are rejuvenated, so that the titer reaches 10<6> or above, air bath shaking culture on host bacteria is carried out at a proper temperature to a logarithmic phase for later use, 100-200 microliters of a LB semi-solid culture medium is added into holes of a 96-hole micro-culture plate respectively, 5-20 microliters of a host bacterium liquid is added after the LBsemi-solid culture medium is solidified, two holes are reserved as negative control, and 5-20 microliters of an LB liquid culture medium is used, 5-20 microliters of diagnostic bacteriophages are added into twp positive control holes, the rest bacteriophages to be detected are respectively added, and after gentle and uniform mixing, the mixture is placed at a proper temperature (18-37 DEG C) for12-24 hours and then is observed. The method provided by the invention can be used for identifying more than 90 bacteriophages or 90 host bacteria each time, the time is short, and the dosage of related reagents and bacteriophages is less.

Description

technical field [0001] The invention relates to a method for high-throughput screening of phage host spectrum, belonging to the technical field of phage main spectrum screening. Background technique [0002] Phage (bacteriophage, phage) is a general term for viruses that infect microorganisms such as bacteria, fungi, algae, actinomycetes, or spirochetes. Some of them can cause lysis of host bacteria, so they are called phages. Phages share some of the properties of viruses: they are tiny. The phage genome contains many genes, but all known phages use the ribosomes of bacteria, various factors required for protein synthesis, various amino acids and energy production systems in bacterial cells to achieve their own growth and proliferation. Once outside the host cell, the phage can neither grow nor replicate. Bacteriophages are widely distributed, and wherever there are bacteria, corresponding phages may exist. In human and animal excrement or polluted well water and river w...

AI Technical Summary

Problems solved by technology

[0003] At present, the determination methods of phage host spectrum at home and abroad are concentrated on direct drop method and double-layer plate method. Whether it is the traditional direct drop method or the classic double-layer plate method, it takes a long time, and the relevant reagents and The waste of phage is large; about 26ml LB medium is needed for the double-layer plate method, and only one host and one phage can be identified

The selection of bacterial strains is mainly host bacteria, related bacteria and non-related bacteria, but a small number of representative strains are selected, which may have omissions, and takes a long time, and the amount of phage and medium used is quite large; therefore It is necessary to find a method for high-throughput screening of phage host spectrum