Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A Copper-resistant Genetic Engineering Bacteria of Acidophilus Thiobacillus Thioxidans and Its Application

A technology of Thiobacillus thiooxidans and genetically engineered bacteria, applied in the directions of bacteria, process efficiency improvement, microorganism-based methods, etc., can solve the problems of environmental hazards, easy loss of plasmids, unfavorable large-scale application of genetically engineered bacteria, etc. Application-wide effects

Active Publication Date: 2016-07-06
SHANDONG UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, in the genus Acidithiobacillus sp., researchers only improve or increase some biological characteristics of the genus by introducing recombinant plasmids; however, the genetic engineering bacteria constructed in the form of plasmids have poor stability and are Under the pressure of no antibiotics, the plasmids in engineering bacteria are easy to lose, which reduces or even loses certain biological characteristics in engineering bacteria. Therefore, in order to improve the stability of plasmids in engineering bacteria, antibiotics must be added to their living environment; acidophilic Thiobacillus is mainly used in bioleaching, heavy metal pollution control and other fields. Adding antibiotics to the above environment will cause harm to the environment on the one hand, and on the other hand, expensive antibiotics will undoubtedly increase industrial costs, which is not conducive to the development of this genetically engineered bacteria. large-scale application of

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A Copper-resistant Genetic Engineering Bacteria of Acidophilus Thiobacillus Thioxidans and Its Application
  • A Copper-resistant Genetic Engineering Bacteria of Acidophilus Thiobacillus Thioxidans and Its Application
  • A Copper-resistant Genetic Engineering Bacteria of Acidophilus Thiobacillus Thioxidans and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Anti-copper gene copB Af Integrated plasmid pZK19-copB Af construction and identification of

[0029] 1. Construction of the plasmid backbone pZK19-IngUH-IngDH for A. thiooxidansATCC19377 gene integration

[0030] On the genome of A.thiooxidansATCC19377, the spacer region between two genes with opposite transcription directions was selected as the insertion site of the exogenous gene; primers were designed to amplify a fragment about 1.0 kb upstream and downstream of the insertion site as the homologous arm of homologous recombination :

[0031] IngUHF: 5'-CTGTCGACGGGGATAGATGACATTC-3'

[0032] IngUHR: 5'-GTAAGCTTGCCTGATCAGCTCACGGT-3'

[0033] IngDHF: 5'-TTATGGATCCGGCTTCAGGCATCGCCGT-3'

[0034] IngDHR: 5'-GGCGCTCTAGACTCTTAATGGTCCTATTCCGAG-3'

[0035] A SalI restriction site was added to the 5' end of the primer IngUHF, a HindIII restriction site was added to the 5' end of the primer IngUHR, a BamHI restriction site was added to the 5' end of the primer I...

Embodiment 2

[0047] Example 2: Construction and Identification of Acidophilus Thiobacillus Thioxidans Copper-Resistant Genetic Engineering Bacteria

[0048] 1. Build copB Af Gene-integrated A.thiooxidans single-crossover strain

[0049] The mid-log phase Escherichia coli S17-1λpir (pZK19-copB Af ) as the donor bacteria and acidophilic thiothiooxidans ATCC19377 in the late logarithmic phase as the recipient bacteria, and mixed according to the ratio of donor bacteria: acceptor bacteria 1:1.5, spread on the Starkey-Na 2 S 2 o 3 On the 0.22um filter membrane of the junction medium plate, culture at 30°C for 7 days; use Starkey-Na 2 S 2 o 3 Inorganic salt solution to elute the bacteria on the filter membrane, and apply to Starkey-Na 2 S 2 o 3 On a solid plate, culture at 30°C for 12 days; then add 1.0 μL of 12 mg / mL sterile X-gal solution dropwise to the above-mentioned single colony, place it at 30°C for 30 minutes, pick a blue colony for PCR amplification and agarose Gel electropho...

Embodiment 3

[0082] Example 3: A. thiooxidanscop B Af (CGMCCNo.9286) Application of Genetic Engineering Bacteria in Bioleaching

[0083] 1.A.thiooxidanscopB Af (CGMCCNo.9286) Growth assay in copper-containing medium

[0084] Acidithiobacillusthiooxidans copB Af Strains in Starkey-S 0 In liquid culture medium, shake the flask at 30°C and 180rpm to the late logarithmic period (about 5-7 days), according to the bacterial concentration of 10 6 ~10 7 A.thiooxidanscopB A.f Inoculate to 100mL containing 0~100mM CuSO 4 Starkey-S 0 In liquid culture medium, shake flask culture at 30°C and 180rpm; take 1mL bacterial suspension from each bottle of bacteria for OD determination on 2 or 3 days during the culture period 600 , and plot A.thiooxidanscopB Af Growth curves under different copper ion concentrations (see figure 2 ), determine A.thiooxidanscopB Af The copper ion MIC of the strain. by A.thiooxidanscopB Af The growth curve of the strain under different copper ion concentrations can...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses copper-resistant genetic engineering bacteria of acidithiobacillus ferroxidans with stable inheritance. The bacterial strain is preserved in China General Microbiological Culture Collection Center on June 9, 2014 with the preservation number of CGMCC No.9286. The engineering bacteria disclosed by the invention are obtained by integrating a copBAf gene of the acidithiobacillus ferroxidans on a genome of the acidithiobacillus ferroxidans. The invention also discloses application of the engineering bacteria in bioleaching. The experiment proves that the copper-resistant genetic engineering bacteria of the acidithiobacillus ferroxidans disclosed by the invention have higher copper resistance characteristics than wild acidithiobacillus ferroxidans, and are stable in inheritance, indicating that the copper-resistant genetic engineering bacteria have great application value in the field of bioleaching of copper ore.

Description

technical field [0001] The invention relates to a strain of acidophilic Thiobacillus thiooxidans and its application, in particular to a strain of acidophilic Thiobacillus thiooxidans capable of stably inheriting copper-resistant engineering bacteria and its application in bioleaching. Background technique [0002] Acidithiobacillus thiooxidans (A.thiooxidans) plays a very important role in the fields of bioleaching, heavy metal pollution control and biological desulfurization, but in the process of bioleaching and heavy metal pollution control, especially in the biological During the leaching process, the copper ion concentration is often very high, generally 30-90mM, and even as high as 300mM in some leaching environments, while the minimum inhibitory concentration (MIC) of copper ions in A.thiooxidans is usually lower than the copper ion concentration in the above-mentioned environment, For example, the copper ion MIC of A.thiooxidansATCC19377 preserved in this experiment...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/74C22B3/18C12R1/01
CPCY02P10/20
Inventor 刘相梅文晴林建群
Owner SHANDONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com