Degenerate primer for rapidly screening and identifying microcystic toxin degrading bacteria and application of degenerate primer

Abstract

The invention discloses a degenerate primer for rapidly screening and identifying microcystic toxin degrading bacteria and application of the degenerate primer. The invention specifically discloses a degenerate primer pair composed of a primer mlrA-354F and a primer mlrA-785R, and a method for rapidly screening and identifying microcystic toxin degrading bacteria. According to the invention, all disclosed mlrA gene sequences are collected from an NCBI database, BioEdit software is utilized to carry out multi-sequence comparison, a degenerate primer pair is designed according to a gene conserved region, DNA of a to-be-detected bacterial strain is taken as a template to carry out PCR amplification and sequencing, a sequencing result is compared with a known sequence, and the MC degradation capability of the bacterial strain with a positive result is detected, to verify the effectiveness of the PCR screening method. All the obtained positive strains can effectively degrade MC, which shows that the identification method provided by the invention has very high accuracy, and can effectively shorten the screening period of degrading bacteria and greatly save the screening cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a degenerate primer for rapid screening and identification of microcystin-degrading bacteria and its application, in particular to the design of a degenerate primer, and the rapid screening and identification of microcystins using the degenerate primer Methods and kits for algal toxin degrading bacteria. Background technique [0002] With the aggravation of eutrophication in water bodies, cyanobacteria blooms occur frequently in aquatic ecosystems around the world, and the release of secondary metabolites-algae toxins caused by them has become a public environmental problem that seriously threatens the ecological environment and human health. Most algal toxins are water-soluble and directly pollute water bodies. Among them, microcystins (microcystins, MC) is the highest detection rate in the cyanobacteria bloom pollution, the most serious algal toxin species. MC can specifically bind...

AI Technical Summary

Problems solved by technology

In general, the successful screening of MC-degrading bacteria needs to detect a large number of strains to be tested, and the workload required to determine their degradation rates one by one is relatively large, and this routine detection method relies on the use of expensive MC pure products (commercially available) MC pure product can reach thousands of yuan per milligram) as the degradation substrate, and the quantitative detection of MC requires liquid chromatography technology, resulting in higher detection cost and longer detection cycle of MC

Therefore, it is urgent to develop a new screening method for MC-degrading bacteria to solve the shortcomings of traditional screening methods.

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