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Heavy metal ion adsorption system, host bacteria thereof and heavy metal ion recovery method

A heavy metal ion and adsorption system technology, applied in the field of genetic engineering, can solve the problems of heavy metal ion recovery selectivity, unsatisfactory effect, and environmental pollution, and achieve the effect of convenient bacterial cell recovery, low cost, and high purity

Inactive Publication Date: 2017-11-07
SHENZHEN Y GENE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, to provide a heavy metal ion adsorption system and its host bacteria, and a method for recovering heavy metal ions, aiming at solving the problem of poor selectivity, low purity and unsatisfactory effect of existing heavy metal ion recovery. and technical issues that pollute the environment

Method used

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  • Heavy metal ion adsorption system, host bacteria thereof and heavy metal ion recovery method
  • Heavy metal ion adsorption system, host bacteria thereof and heavy metal ion recovery method
  • Heavy metal ion adsorption system, host bacteria thereof and heavy metal ion recovery 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 heavy metal ion adsorption system, comprising the following steps:

[0037] S011: Amplify the Bacillus subtilis spore coat protein CotC DNA sequence from the Bacillus subtilis genome;

[0038] S012: Amplifying the heavy metal ion binding protein DNA sequence from the strain genome expressing the heavy metal ion binding protein;

[0039]S013: connecting the promoter, the CotC DNA sequence of the Bacillus subtilis spore coat protein and the DNA sequence of the heavy metal ion binding protein in sequence.

[0040] In one embodiment, the primers for amplifying the CotC DNA sequence of the Bacillus subtilis spore coat protein are shown in SEQ ID NO: 1, SEQ ID NO: 2; in another embodiment, the promoter can be Pcot or Pveg, And the primers for amplifying Pcot are shown in SEQ ID NO:16, SEQ ID NO:17, the primers for amplifying Pveg are shown in SEQ ID NO:19, SEQ ID...

Embodiment 1

[0048] Example 1 Construction and Identification of Expression Plasmid of Heavy Metal Ion Detection System

[0049] 1. Amplification of heavy metal ion adsorption gene fragments in the detection system

[0050] Genomic DNA from Salmonella typhimurium using specific primers (SEQ ID NO:4)(SEQ ID NO:5)(SEQ ID NO:7)(SEQ ID NO:8)(SEQ ID NO:10)(SEQ ID NO:11) (Cited in NCBI: NC_003197 ) and the Cusperophilus genomic DNA (reference location in NCBI: NC_007973 ) to amplify the DNA sequence encoding the heavy metal ion adsorption gene and the DNA sequence of the artificially synthesized SUP gene (SEQ ID NO: 6) (SEQ ID NO: 9) (SEQ ID NO: 12), the reaction conditions are as follows.

[0051] PCR reaction system (50μl):

[0052]

[0053] PCR conditions:

[0054] Step 1: 95°C, 3 minutes

[0055] Step 2: 95°C, 1 minute

[0056] Step 3: 60°C, 2 minutes

[0057] Step 4: 72°C, 4 minutes

[0058] Step 5: Repeat steps 2 to 4, 29 cycles

[0059] Step 6: 72°C, 10 minutes

[0060] ...

Embodiment 2

[0135] Example 2 Construction and Identification of Expression Plasmid of Heavy Metal Ion Adsorption System

[0136] 1. Amplification of the dedicated promoter fragments Pcot and Pveg in the adsorption system

[0137] According to the target DNA sequence to be amplified and the principle of base pairing, design specific primers (SEQ ID NO: 16) (SEQ ID NO: 17) (SEQ ID NO: 19) (SEQ ID NO: 20), from our laboratory The Pcot sequence (SEQ ID NO: 18) and the Pveg sequence (SEQ ID NO: 21) of the promoter of the heavy metal ion-regulated gene were amplified from the Bacillus subtilis genome, and the reaction conditions were as follows.

[0138] PCR reaction system (50μl):

[0139]

[0140] PCR reaction conditions:

[0141] Step 1: 95°C, 2 minutes

[0142] Step 2: 95°C, 0.5 minutes

[0143] Step 3: 63°C, 0.5 minutes

[0144] Step 4: 72°C, 0.5 minutes

[0145] Step 5: Repeat steps 2 to 4, 29 cycles

[0146] Step 6: 72°C, 10 minutes

[0147] Step 7: keep warm at 4°C.

[01...

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Abstract

The invention belongs to the technical field of genetic engineering, and particularly relates to a heavy metal ion adsorption system, host bacteria thereof and a heavy metal ion recovery method. The heavy metal ion adsorption system comprises a bacillus subtilis spore coat protein CotC DNA sequence and a heavy metal ion binding protein DNA sequence which are connected with each other, and the bacillus subtilis spore coat protein CotC DNA sequence is connected with a promoter. A heavy metal ion attachment protein surface display system includes bacillus subtilis spore coat protein CotC and heavy metal ion binding protein which are expressed by the heavy metal ion adsorption system provided by the invention. The recovery method provided by the invention can realize the enrichment and recovery of heavy metal ions in industrial waste water through a biological means, and can overcome the disadvantages of the prior art without causing secondary pollution and the environment-friendly effect is realized.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a heavy metal ion adsorption system, host bacteria, and a method for recovering heavy metal ions. Background technique [0002] In recent years, with the rapid development of my country's economic construction, the mining, smelting, processing and commercial manufacturing activities of heavy metals are increasing, causing many heavy metals (such as lead, mercury, cadmium, etc.) A series of serious pollution incidents have caused great harm to the ecological environment. Acute poisoning caused by heavy metals and various chronic human diseases such as Alzheimer's disease are also threatening human health at all times. On the other hand, with the rapid development of science and technology, those heavy metals with stable chemical properties and excellent physical properties, such as gold, silver, copper, platinum, palladium, etc., are widely used in electron...

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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