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Construction and application of broad spectrum organic phosphorus pesticide degradation genetic engineering strain

A technology of genetically engineered strains, genetically engineered bacteria, applied in the direction of genetic engineering, application, plant genetic improvement, etc.

Inactive Publication Date: 2012-01-11
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The random insertion vector pUT mini-Tn5 has been used as an insertion tool to construct engineering strains, but there is no transposon random insertion vector without exogenous resistance genes in the world

Method used

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  • Construction and application of broad spectrum organic phosphorus pesticide degradation genetic engineering strain
  • Construction and application of broad spectrum organic phosphorus pesticide degradation genetic engineering strain
  • Construction and application of broad spectrum organic phosphorus pesticide degradation genetic engineering strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0028] 1. Construction of genetically engineered strains for degrading broad-spectrum organophosphorus pesticides

[0029] 1) Organophosphorus pesticide hydrolase genes mpd and oph are inserted into the vector pUTTnsTet

[0030] The vector (pUTTnsTet) (Rong Li, Guangli Wang, Bin Shen, Rong Wang, Yao Song, Shunpeng Li, Jiandong Jiang, Random transposon vectors pUTTns for the markerless integration of exogenous genesinto gram-negative eubacteria chromosomes, Journal of Microbiological Methods, 2009(79): 220~226) Escherichia coli was cultured in LB medium to the late logarithmic stage, the plasmid was extracted, and the vector pUTTnsTet was digested with restriction enzyme ApaI to recover linear fragments; degraded from organophosphorus pesticides Psedudomonas putidaDLL-1 (Liu Zhi, Sun Jianchun, Li Shunpeng. Isolation, identification and degradation of methyl parathion-degrading bacteria DLL-1. Journal of Applied and Environmental Biology, 1999, 147-151) The mpd gene was amplified (p...

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Abstract

The invention discloses construction and application of a broad spectrum organic phosphorus pesticide degradation genetic engineering strain, and belongs to the field of biological engineering technology. The engineering strain 1gmp is the engineering strain paracoccus sp. 1gmp for degrading the organic phosphorus pesticide in a broad spectrum, which is constructed by sequentially inserting two organic phosphorus hydrolase genes mpd and oph with different substrate degradation spectrums into the chromosome of dimethoate high-efficiency degradation strain paracoccus sp. L3 by using random integration of a serious of vectors pUTTns and deleting a resistant gene by counter-selection marking; and degradation spectrum measurement shows that the engineering strain can efficiently degrade 17 organic phosphorus pesticides generally used in China at present. The engineering strain has the advantages of genetic stability, biological safety and great potential application prospect.

Description

1. Technical Field [0001] The invention relates to the construction and application of a broad-spectrum organophosphorus pesticide degradation genetic engineering bacteria, and belongs to the technical field of bioengineering. 2. Background technology [0002] Organophosphorus pesticides can inhibit the activity of acetylcholinesterase, accumulate acetylcholine (a neurotransmitter) in body tissues, and cause a series of poisoning symptoms such as certain neurological disorders. In severe cases, it can paralyze the nerves and cause death. The use of organophosphorus pesticides poses a potential threat to human health or the ecological environment. Many countries have identified organophosphorus pesticides such as dimethoate, dichlorvos, and parathion as environmental priority pollutants. In the early 1980s, the United States began to investigate the problem of pesticides contaminated groundwater caused by poisoning accidents. In some areas, more than 50 pesticide residues were det...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63A62D101/04C12N15/55C12N1/21C12R1/01A62D101/26A62D3/02
Inventor 蒋建东李荣李顺鹏王融宋瑶朱建春
Owner NANJING AGRICULTURAL UNIVERSITY