Application of chlamydomonas reinhardtii VPS9 gene in regulating and controlling cadmium resistance of chlamydomonas reinhardtii

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

Inactive Publication Date: 2018-03-06
JIANGHAN UNIVERSITY
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  • Description
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  • Application Information

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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 VPS9 gene in regulating and controlling cadmium resistance of chlamydomonas reinhardtii
  • Application of chlamydomonas reinhardtii VPS9 gene in regulating and controlling cadmium resistance of chlamydomonas reinhardtii
  • Application of chlamydomonas reinhardtii VPS9 gene in regulating and controlling cadmium resistance of chlamydomonas reinhardtii

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Embodiment 1

[0035] 1. Obtaining Chlamydomonas reinhardtii transformants by electroporation

[0036] (1) Prepare the aphⅧ fragment

[0037] The PJMG-aphⅧ plasmid containing the 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 electrophoresis results after enzyme digestion are as follows figure 1 As shown, M is BM5000 DNA Marker, 1 is the EcoRI digest product of PJMG-aphⅧ plasmid, 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.

[0038] (2) Chlamydomonas cells prepared for...

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Abstract

The invention discloses application of a chlamydomonas reinhardtii VPS9 gene in regulating and controlling cadmium resistance of chlamydomonas reinhardtii, and belongs to the technical field of biologics and the field of sewage improvement. According to the application, an aphVIII fragment (SEQ ID NO:3) with a paromomycin resistance gene is randomly inserted into chlamydomonas, then a chlamydomonas reinhardtii mutant alga strain which is remarkably sensitive to cadmium is cultured, and further identification shows that the cadmium sensitivity of the mutant alga strain is caused since the aphVIII fragment is inserted into a VPS9 gene (a CDS (Cadmium Sulfide) sequence is shown in SEQ ID NO:1 in the specification). By reducing expression of the chlamydomonas reinhardtii VPS9 gene, deactivating the VPS9 gene or by deleting the VPS9 gene, the cadmium resistance of the chlamydomonas reinhardtii can be degraded; a cadmium-sensitive chlamydomonas reinhardtii strain obtained by means of geneticengineering can be applied to monitoring and detection on cadmium polluted water areas, and novel ideas and biological materials can be provided for sewage treatment by using a biological method.

Description

technical field [0001] The invention relates to the fields of biotechnology and sewage treatment, in particular to the application of the Chlamydomonas reinhardtii VPS9 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 i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N1/13C07K14/405C12R1/89
Inventor 李丽丽章伟雄李甜甜方治伟刘艳玲魏志新何苗李论
Owner JIANGHAN UNIVERSITY
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