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Construction and application of engineering bacterium LZ-17

A technology of engineering bacteria and application methods, applied in the fields of application, genetic engineering, bacteria, etc., can solve the problems of laborious, human error, time-consuming, etc., and achieve the effect of shortening the bacterial culture time, eliminating human error, and high sensitivity

Inactive Publication Date: 2003-10-15
EAST CHINA NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the background technology is that plate counting can only be performed manually, which is time-consuming, labor-intensive, prone to human error, and cannot be automated.
In addition, the test results must wait for 48 hours before they can be obtained. In the face of the situation where there are many samples to be tested and the test results are required to be obtained as soon as possible, the background technology is often helpless

Method used

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  • Construction and application of engineering bacterium LZ-17
  • Construction and application of engineering bacterium LZ-17

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Application of engineering bacteria LZ-17 to determine the mutagenicity of 4-nitro-o-phenylenediamine

[0042] Except that the tested sample is 4-nitro-o-phenylenediamine, that is, NPD, the operations of the first, second, third, fourth and fifth steps are all identical to those of the corresponding steps of the application method of the present invention. The average value of 8 MR values ​​measured in the fifth step is 2.2, and it is determined that the mutagenicity of 4-nitro-o-phenylenediamine, that is, NPD, is positive.

Embodiment 2

[0043] Example 2 Determining the Mutagenicity of Diaminofluorene Using Engineering Bacteria LZ-17

[0044] Except that the tested sample is diaminofluorene, that is, 2-AF, the operations of the first, second, third, fourth, fifth, sixth and seventh steps are all identical to those of the corresponding steps of the application method of the present invention. In the fifth step, the average value of the eight MR values ​​measured is less than 2, and the sixth and seventh steps are continued. In the seventh step, the average value of 8 MR values ​​is 2.56, and it is determined that the mutagenicity of diaminofluorene, ie 2-AF, is positive.

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Abstract

The present invention relates to the construction and application of engineering bacterium LZ-17, and belongs to the field of bioengineering and gene engineering technology. By means of molecular cloning technology, the luminous gene of Qinghai vibrio is introduced into Salmonella typhimurium TA97 to constitute engineering bacterium LZ-17. The engineering bacterium can produce bacterioleuciferinase, can glow with visible light of 440-640 nm wavelength after decanol is added, and has the Ames test relevant characters the same as Salmonella typhimurium TA97. The engineering bacterium is applied in Ames test as testee life form for determining the number of back varying bacterium. The engineering bacterium is especially suitable for detection of mutagenicity of the testee sample.

Description

technical field [0001] The invention relates to the construction and application of engineering bacterium LZ-17, that is, the construction method and application method of engineering bacterium LZ-17, belonging to the technical field of bioengineering or genetic engineering. Background technique [0002] The Ames test is a method to detect the mutagenicity of the tested samples. The method adopts Salmonella typhimurium (Salmonella typhimurium) TA97 as the test organism, including the following basic operations: 1. Bacteria enrichment: cultivate Salmonella typhimurium TA97 at 37° C. for 12 hours with a nutrient broth medium shake flask, and then take it out from the shake flask 0.1ml (add 10% S-9 solution to the sample that needs to be activated), put it into the basic medium together with the sample to be tested, mix well, pour it into the plate with the bottom medium on the bottom of the plate, put the plate in a temperature of 37 ℃ 2. Plate countin...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/09C12N15/31C12Q1/02
Inventor 朱文杰杨晔晔景奉香赵曙霞
Owner EAST CHINA NORMAL UNIVERSITY
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