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Uranium-related gene segments of pokeweed and acquiring method and application

A technology of gene fragments and pokeweed, applied in the fields of molecular biology and environmental monitoring

Inactive Publication Date: 2010-08-04
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, this method has not been reported in China

Method used

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  • Uranium-related gene segments of pokeweed and acquiring method and application
  • Uranium-related gene segments of pokeweed and acquiring method and application
  • Uranium-related gene segments of pokeweed and acquiring method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Take the pokeweed root tissue from the uranium tailings area and the non-polluted control area, and store it in liquid nitrogen at -196°C until use;

[0033] 2. Extract total RNA from pokeweed root tissue, such as figure 1

[0034] (1) Get pre-cooled RNA extraction buffer (4mol / L guanidine isothiocyanate, 25mmol / L sodium citrate (pH7.0), 0.5% (W / V) sodium lauryl sarcosine, 4 % (V / V) β-mercaptoethanol (added before use)) 10mL was added to a 50mL centrifuge tube without RNase, and placed on ice;

[0035] (2) Weigh 4g of pokeweed root system and grind it fully in a mortar with excess liquid nitrogen, transfer the pokeweed root system powder into a centrifuge tube with a pre-cooled medicine spoon, shake vigorously for 10 minutes, and stand on ice for 5 minutes ;

[0036] (3) Add 1mL 2mol / L sodium acetate (pH 4.0) and shake well;

[0037] (4) Add 10 mL of water-saturated phenol (pH 4.5) and shake well;

[0038] (5) Add 2 mL of chloroform:isoamyl alcohol (24:1), shake...

Embodiment 2

[0155] 1. The steps are the same as in Example 1;

[0156] 2. The steps are the same as in Embodiment 1 and 2;

[0157] 3. Step is the same as embodiment one 3;

[0158] 4. Step is the same as embodiment one 4;

[0159] 5. The steps are the same as in Embodiment 1 and 5;

[0160] 6. The steps are the same as in Example 1; 6;

[0161] 7. Design primers based on the sequences at both ends of the specifically expressed gene fragment;

[0162] 8. Do RT-PCR to detect pokeweed cDNA in the environment.

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Abstract

The invention discloses uranium-related gene segments of pokeweed. The method for acquiring the gene segments comprises the following steps: extracting total RNA from root tissues of pokeweed in uranium tailings region and non-polluted contrast region, synthesizing a first chain and a second chain of cDNA by enzymatic reaction reverse transcription, carrying out suppression subtractive hybridization to separate gene segments with specific expression under heavy metal uranium stress, using the gene segments as probes and enabling the gene segments and RNA extracted from pokeweed in detected environment to hybridize, or using sequences of two ends of the gene segments with specific expression as primers to carry out reverse transcription-polymerase chain reaction to detect the cDNA of pokeweed in the detected environment. In the invention, gene with specific expression induced by uranium in pokeweed is used as a probe to detect uranium pollution, solving the problem of inaccurate detection of uranium pollution in surrounding environment of uranium-polluted areas.

Description

technical field [0001] The present invention belongs to the fields of molecular biology and environmental monitoring, and specifically discloses a gene fragment related to uranium in pokeweed and a method for obtaining the gene fragment, and further relates to the use of the gene fragment in detecting uranium pollution in the environment application. Background technique [0002] The process of uranium mining and nuclear energy utilization is an important way for uranium to be enriched and polluted in the environment. Uranium tailings and waste rock contain uranium and all its daughters, and its radionuclide content is 2 to 3 orders of magnitude higher than the background. Improper accumulation or disposal of these wastes can result in uranium being dispersed to the soil surface through runoff, into the air through wind erosion, or into groundwater through leaching. In addition, the uranium ore smelting process will continuously discharge a large amount of radioactive wast...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12P19/34C12Q1/68
Inventor 高利臣肖璐冯涛
Owner HUNAN UNIV OF SCI & TECH
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