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Gold ion detection and adsorption system, host bacteria thereof and gold ion recycling method

An adsorption system and gold ion technology, applied in the field of genetic engineering, can solve the problems of poor gold ion recovery selectivity, unsatisfactory effect, and environmental pollution, and achieve the effect of convenient bacteria recovery, low cost, and environmental friendliness

Inactive Publication Date: 2017-11-10
SHENZHEN Y GENE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, to provide a gold ion detection and adsorption system and its host bacteria, and a gold ion recovery method, aiming to solve the problem of poor selectivity, low purity and ineffective effect of existing gold ion recovery. ideal, and the technical problem of polluting the environment

Method used

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  • Gold ion detection and adsorption system, host bacteria thereof and gold ion recycling method
  • Gold ion detection and adsorption system, host bacteria thereof and gold ion recycling method
  • Gold ion detection and adsorption system, host bacteria thereof and gold ion recycling method

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preparation example Construction

[0036] In yet another aspect, an embodiment of the present invention provides a method for preparing the above-mentioned gold ion detection and adsorption system, which is characterized in that it includes the following steps:

[0037] S011: Amplify the Bacillus subtilis spore coat protein CotC DNA sequence from the Bacillus subtilis genome, and amplify the gold ion-binding protein GolB DNA sequence from the Salmonella typhimurium genome;

[0038] S012: Using the above-mentioned Bacillus subtilis spore coat protein CotC DNA sequence and the gold ion binding protein GolB DNA sequence as templates, amplify the CotC-GolB fusion protein DNA sequence;

[0039] S013: Amplify the fluorescent protein DNA sequence from the Escherichia coli expression vector genome;

[0040] S014: connecting the first promoter, the fluorescent protein DNA sequence, the second promoter, and the CotC-GolB fusion protein DNA sequence sequentially.

[0041] In one embodiment, primers for amplifying the DNA...

Embodiment 1

[0050] Example 1 Construction and Identification of Expression Plasmid of Gold Ion Detection System

[0051] 1. Gold ion-induced gene in the detection system: promoter P golS -GolS-P golB amplify

[0052] Using specific primers pGolts1 (SEQ ID NO: 8) and pGolts2 (SEQ ID NO: 9) from Salmonella typhimurium genomic DNA (reference position in NCBI: NC_003197 ) to amplify the DNA sequence (SEQ ID NO: 10) encoding the gold-induced gene, and the reaction conditions are as follows. The gold ion-induced gene (Gold inductive gene) of this embodiment is the promoter P golS -GolS-P golB GolS, which can express GolS protein, GolS protein usually acts as a repressor, and only when there is gold ion, it will be released from the promoter P golB The DNA sequence is shed, triggering the expression of downstream genes.

[0053] PCR reaction system (50μl):

[0054]

[0055] PCR reaction conditions:

[0056] Step 1: 95°C, 3 minutes

[0057] Step 2: 95°C, 1 minute

[0058] Step 3: ...

Embodiment 2

[0122] Example 2 Construction and Identification of Expression Plasmid of Gold Ion Adsorption System

[0123] 1. Promoter P in the adsorption system golB fragment amplification

[0124] According to the target DNA sequence to be amplified and the principle of base pairing, design specific primers P golB 1 (SEQ ID NO: 17) and P golB 2 (SEQ ID NO: 18), the promoter P of the gold ion-regulated gene was amplified from all E. golB The sequence (SEQ ID NO: 19), the reaction conditions are as follows.

[0125] PCR reaction system (50μl):

[0126]

[0127] PCR reaction conditions:

[0128] Step 1: 95°C, 2 minutes

[0129] Step 2: 95°C, 0.5 minutes

[0130] Step 3: 63°C, 0.5 minutes

[0131] Step 4: 72°C, 0.5 minutes

[0132] Step 5: Repeat steps 2 to 4, 29 cycles

[0133] Step 6: 72°C, 10 minutes

[0134] Step 7: keep warm at 4°C.

[0135] Recover the PCR product for later use.

[0136] 2. Amplification of gold ion adsorption gene fragments in the adsorption system ...

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Abstract

The invention belongs to the technical field of genetic engineering, and particularly relates to a gold ion detection and adsorption system, host bacteria thereof and a gold ion recycling method. The gold ion detection and adsorption system comprises a detection unit and an adsorption unit connected with the detection unit, the detection unit comprises a fluorescent protein DNA sequence, and the fluorescent protein DNA sequence is connected with a first promoter; the adsorption unit comprises a bacillus subtilis spore coat protein CotC DNA sequence and a gold ion binding protein GolB DNA sequence which are mutually connected, and the bacillus subtilis spore coat protein CotC DNA sequence is connected with a second promoter; the detection unit is located at the upstream of the adsorption unit. According to the gold ion detection and adsorption system, host bacteria thereof and the gold ion recycling method, detection and recyclable enrichment and recycling of heavy metal gold in industrial wastewater can be achieved through a biological means, the disadvantages of the prior art can be overcome, secondary contamination cannot be caused, and the gold ion detection and adsorption system is environmentally friendly.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a gold ion detection and adsorption system, its host bacteria, and a gold ion recovery method. Background technique [0002] Gold is a precious metal discovered and utilized by human beings earlier. Due to its scarcity of resources and unique physical and chemical properties, gold has been regarded as the head of hardware since ancient times, and has the title of "King of Metals". Gold is a rare metal with excellent chemical, physical and electronic properties among heavy metals. It has high stability and its application fields are very wide. Because gold has very stable physical and chemical properties, such as corrosion resistance, electrical conductivity, thermal conductivity and stability, it can be widely used in many modern high-tech, and gold ions can be used as inhibitors of white blood cells and other immune cells in the immune system . In addit...

Claims

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Application Information

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IPC IPC(8): C12N15/62C07K19/00C12N1/21C12N15/70C02F3/34C22B3/18C22B11/00C12R1/19C02F101/20
CPCC02F3/34C02F2101/20C07K14/195C07K14/255C07K14/32C07K2319/60C12N15/70C22B3/18C22B11/04Y02P10/20
Inventor 赵劲魏炜孙培卿崔汉立刘祥志
Owner SHENZHEN Y GENE BIOTECH CO LTD
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