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Acidithiobacillus caldus gene engineering strain and applications thereof

A technology of thermophilic Thiobacillus and genetic engineering bacteria, applied in the field of genetic engineering, can solve problems such as improving the sulfur oxidation ability of acidophilic Thiobacillus thermophilic, achieve the effects of improved transcription level, enhanced ability to oxidize sulfur, and wide application prospects

Inactive Publication Date: 2014-02-12
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, there is no relevant report on improving the sulfur oxidation ability of acidophilus thermophilic Thiobacillus acidophilus in a transgenic way

Method used

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  • Acidithiobacillus caldus gene engineering strain and applications thereof
  • Acidithiobacillus caldus gene engineering strain and applications thereof
  • Acidithiobacillus caldus gene engineering strain and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0056] (1) Strain selection:

[0057] Acidithiobacillus caldus MTH-04; Escherichia coli DH5α.

[0058] (2) Construction of the cloning vector pUC19-sor:

[0059] The SOR protein coding gene sor was cloned from the chromosomal DNA of Thiobacillus acidothermophilus MTH-04, and the extraction method of the chromosomal DNA of Thiobacillus acidothermophilus MTH-04 can be found in Tiangen Bacteria Genomic DNA Extraction Kit (purchased from Tiangen Biochemical Technology Co., Ltd. Company) instructions, the nucleotide sequences of the primers used are as follows:

[0060] sor upstream primer sor Forward:

[0061] 5'-CCG GAATTC TGGAGGCAATGTGGACAAAAATCCT-3'

[0062] EcoR Ⅰ

[0063] sor downstream primer sor Reverse:

[0064] 5'-TGC TCTAGA CTCA GTGATGATGATGATGATG CACAAGTTTGCGCCGCCAT-3'

[0065] Xba Ⅰ 6×His tag

[0066] PCR reaction system: 5×Buffer: 10 μl; dNTP: 4 μl; DNA template: 2 μl; forward and reverse primers 0.5 μl each; PrimeStar enzyme (TaKaRa company)...

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Abstract

The invention relates to an acidithiobacillus caldus gene engineering strain and applications thereof. The acidithiobacillus caldus MTH-04(pJRD215-tac-sor) gene engineering strain is preserved in the general microbiology centre of China Committee for Culture Collection of Microorganisms on June 28, 2013, address: Institute of Microbiology Chinese Academy of Sciences, NO.3, NO.1 yard, beichen west road, Beijing chaoyang district. The strain preservation number: CGMCC NO.7388. Compared with the acidithiobacillus caldus MTH-04, the acidithiobacillus caldus MTH-04(pJRD215-tac-sor) gene engineering strain has the advantages that the capability of sulfur oxide is enhanced, and the bacteria biomass, sulfur and oxygen reductase enzyme activity and the transcriptional level of key enzyme of a sulfur oxidation system are increased.

Description

technical field [0001] The invention relates to a strain of acidophilic Thiobacillus thermophilic genetic engineering bacteria and its application, in particular to a genetic engineering bacteria strain of acidophilic Thiobacillus thermophilicus with higher sulfur oxidation ability and its application, which belongs to the technical field of genetic engineering. Background technique [0002] With the advantages of improving resource utilization and reducing environmental pollution, biometallurgy technology has shown broad application prospects in the field of rare metal recovery. However, the low oxidation rate and long oxidation cycle of autotrophic bacteria used in biometallurgy have always been the bottleneck of the application of this technology. . It is an urgent problem to be solved to genetically modify autotrophic bacteria to construct engineering bacteria with fast growth rate and strong oxidation ability, and to improve their application efficiency. [0003] Acidi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C22B3/18B01D53/84B01D53/48C02F11/02C02F3/34C12R1/01
CPCY02A50/20Y02P10/20
Inventor 庞昕李良林建群崔爽刘相梅林建强
Owner SHANDONG UNIV
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