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Gold ion adsorption protein and method for enriching and recovering gold ion

A gold ion and protein technology, which is applied in the field of bioengineering, can solve the problems of difficult and efficient selective enrichment and recovery of gold ions, and achieves the effect of convenient operation and low price.

Inactive Publication Date: 2013-02-13
SHENZHEN Y GENE BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing methods are difficult to achieve high-efficiency selective enrichment and recovery of gold ions in industrial wastewater.

Method used

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  • Gold ion adsorption protein and method for enriching and recovering gold ion
  • Gold ion adsorption protein and method for enriching and recovering gold ion
  • Gold ion adsorption protein and method for enriching and recovering gold ion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Construction and identification of embodiment 1OmpA-GolB fusion protein

[0045] 1. Gene Amplification of Escherichia coli Outer Membrane Protein A (OmpA) Fragment

[0046] According to the target DNA sequence to be amplified and the principle of base pairing, design specific primers Ompa1 (SEQ ID NO: 1) and Ompa2 (SEQ ID NO: 2), from Escherichia coli genomic DNA (reference position in NCBI: NC_000913 ) to amplify the DNA sequence (SEQ ID NO: 3) encoding the first to 159th amino acids of Escherichia coli outer membrane protein A (OmpA).

[0047] The reaction conditions are as follows:

[0048] PCR reaction system: 50μl

[0049] wxya 2 O: 36 μl

[0050] 10X Pfu DNA Polymerase Buffer: 5 μl

[0051] 10mM dNTPs: 1μl

[0052] Forward primer Ompa1 1 μl

[0053] Reverse primer Ompa2 1μl

[0054] Pfu DNA polymerase: 1 μl

[0055] DNA template: 5μl

[0056] PCR conditions:

[0057] The first step is 95°C for 2 minutes

[0058] The second step is 95°C for 0.5 minutes...

Embodiment 2

[0116] Example 2 Construction and identification of Antigen43-gold ion-inducible gene (Gold inductive gene with antigen43) fusion protein expression plasmid

[0117] 1. Gold ion-induced gene amplification

[0118] Using specific primers pGolts1 (SEQ ID NO: 8) and pGolb2 (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 inductive gene.

[0119] The reaction conditions are as follows:

[0120] PCR reaction system: 50μl

[0121] wxya 2 O: 36 μl

[0122] 10X Pfu DNA polymerase buffer: 5 μl

[0123] 10m MdNTP: 1 μl

[0124] Forward primer pGolts1 1 μl

[0125] Reverse primer pGolb2 1 μl

[0126] Pfu DNA polymerase: 1 μl

[0127] DNA template: 5μl

[0128] PCR conditions:

[0129] Step 1: 95°C for 3 minutes

[0130] Step 2: 95°C for 1 minute

[0131] Step 3: 60°C for 2 minutes

[0132] Step 4: 72°C for 4 minutes

[0133] The fifth step repeats the second step t...

Embodiment 3

[0147] Example 3 OmpA-GolB fusion protein, Antigen43-gold ion-induced gene (Gold inductive gene with antigen43) fusion protein transfer into host bacteria and culture of host bacteria

[0148] 1. OmpA-GolB fusion protein into host bacteria

[0149] 1) The plasmid used to express the OmpA-GolB fusion protein was introduced into the Escherichia coli cell DH10b by chemical transformation method, by adding ampicillin (50 μg mL -1 ) on LB solid medium at 37°C for 16 hours, from which positive clones were screened out and preserved.

[0150] The heat shock transformation and dilution coating steps of competent cells are as follows:

[0151] ①Put the product of the ligation reaction in the competent cell (DH10b), and mix it evenly by finger flicking;

[0152] ② After 30 minutes of ice bathing, heat shock at 42°C for 95 seconds, and quickly transfer to ice;

[0153] ③ After 2 minutes in ice bath, add 900 μl of preheated LB medium at 37°C to each tube, and incubate at 37°C, 200 rp...

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Abstract

The invention provides gold ion adsorption protein and a method for enriching and recovering gold ions. The gold ion adsorption protein comprises an escherichia coli outer membrane protein A fragment and a gold ion binding protein GolB fragment; or comprises a gold ion induced protein fragment and an antigen 43 fragment. The gold ion adsorption protein and the method for enriching and recovering gold ion disclosed by the invention can overcome disadvantages in the prior art, and cause no secondary pollution; the gold ion adsorption protein can efficiently adsorb gold ions in the presence of various heavy metal ions, is environment-friendly, and cause no effect on the environment; and the gold ion adsorption protein is easy to prepare, low in cost, and convenient for operation.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to the technology of transforming bacteria through gene and protein engineering, and in particular to a method for gold ion adsorption protein and enrichment and recovery of gold ions. Background technique [0002] How to rationally apply heavy metals in nature to human production and life, so as to make them develop in harmony with ecological environment protection and sustainability is a major challenge for scientific and technological workers at present. On the one hand, some heavy metals are extremely toxic to organisms at very low concentrations, and unlike organic compounds, heavy metals are enriched and difficult to degrade in the natural environment. 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, mer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N1/21C02F3/34C12R1/19C12R1/42
Inventor 陈鹏赵劲魏炜陈兴
Owner SHENZHEN Y GENE BIOTECH CO LTD
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