Application of chlamydomonas reinhardtii VMPL1 gene to regulation of cadmium tolerance of chlamydomonas reinhardtii

A rhinestone and tolerance technology, applied in the fields of biology and sewage treatment, can solve the problems of complex operation, high cost, secondary pollution, etc., and achieve the effect of mature technology system and simple method

Inactive Publication Date: 2018-04-13
JIANGHAN UNIVERSITY
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Problems solved by technology

In comparison, traditional physical and chemical methods are often costly and complicated to operate, and are suitable for concentrated treatment of high-concentration heavy metal-contaminated areas. If they are used in large areas, they will easily cause secondary pollution

Method used

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  • Application of chlamydomonas reinhardtii VMPL1 gene to regulation of cadmium tolerance of chlamydomonas reinhardtii
  • Application of chlamydomonas reinhardtii VMPL1 gene to regulation of cadmium tolerance of chlamydomonas reinhardtii
  • Application of chlamydomonas reinhardtii VMPL1 gene to regulation of cadmium tolerance of chlamydomonas reinhardtii

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Embodiment 1

[0036] 1. Obtaining Chlamydomonas reinhardtii transformants by electroporation

[0037] (1) Prepare the aphⅧ fragment

[0038] PJMG-aphⅧ plasmid containing aphⅧ fragment (Zhangfeng Hu, Yinwen Liang, Wei He, Junmin Pan * , Cilia Disassembly with Two Distinct Phases of Regulation, CellReports, 2015, 10(11): 1803-1810) was digested with EcoRI enzyme at 37°C overnight, and the aphⅧ fragment was separated by 1% agarose gel electrophoresis (2118bp, sequence such as SEQ ID NO :3), the result of electrophoresis after enzyme digestion is as follows figure 1 As shown, M is DL2000 DNA Marker, 1 is the pJMG-aphⅧ plasmid EcoRI digestion product, and the band around 2k is the aphⅧ fragment. The aphⅧ fragment was recovered from the gel with Sangon’s Gel Recovery Kit (Cat. No. B518191-0100), and the concentration of the recovered DNA fragment was measured with a Q5000 ultra-micro nucleic acid detector, and stored at -20°C for future use.

[0039] (2) Chlamydomonas cells prepared for electr...

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Abstract

The invention discloses application of chlamydomonas reinhardtii VMPL1 gene to the regulation of the cadmium tolerance of chlamydomonas reinhardtii, and belongs to the fields of biotechnology and environmental pollution control. According to the invention, an aph VIII segment (SEQ ID NO:3) containing paromomycin resistance gene is randomly inserted into chlamydomonas, chlamydomonas reinhardtii mutant strain obviously sensitive to cadmium is obtained, and through further identification, the fact that the cadmium resistance of the mutant strain is caused because the aph VIII segment is insertedinto the VMPL1 gene (CDS sequence is as shown in SEQ ID NO:1) is discovered. By reducing the expression of the chlamydomonas reinhardtii VMPL1 gene to enable the VMPL1 gene to be inactivated or deleted, the VMPL1 gene can improve the cadmium tolerance of chlamydomonas reinhardtii; the chlamydomonas reinhardtii strain with enhanced cadmium tolerance obtained through a gene engineering method can beapplied to the mentoring and control of cadmium polluted waters. According to the invention, a novel thought and a biological material are provided for sewage treatment.

Description

technical field [0001] The invention relates to the fields of biotechnology and sewage treatment, in particular to the application of the Chlamydomonas reinhardtii VMPL1 gene in regulating the cadmium tolerance of the Chlamydomonas reinhardtii. Background technique [0002] With the rapid development of social economy and industrial production, the problem of environmental pollution is becoming more and more serious, especially the problem of heavy metal pollution. There are many sources of heavy metal pollution. The "three wastes" (waste water, waste residue and waste gas) discharged from factories, vehicle exhaust, pesticide residues, and various domestic waste and waste electronic products produced in people's daily activities all contain heavy metals. The "three wastes" Exceeding standards and illegal discharges, as well as random discarding of domestic garbage and waste electronic products, can easily cause a large amount of toxic and harmful heavy metals to accumulate ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N1/13C07K14/405C12Q1/02C02F3/32C12R1/89C02F101/20
CPCC02F3/322C02F2101/20C07K14/405C12Q1/02G01N2333/405
Inventor 李丽丽何苗章伟雄彭海高利芬陈斌田广梅唐宇云胡长峰
Owner JIANGHAN UNIVERSITY
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