Application of Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating cadmium tolerance of Chlamydomonas reinhardtii

A cre01.g046237.t1.1, Rhine clothing technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of high cost and complicated operation, achieve simple method, mature technical system, significant resistance to heavy metal cadmium Effect

Inactive Publication Date: 2018-03-23
JIANGHAN UNIVERSITY
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In comparison, traditional physical and chemical methods often have the characteristics of high cost and complicat

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
  • Application of Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating cadmium tolerance of Chlamydomonas reinhardtii
  • Application of Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating cadmium tolerance of Chlamydomonas reinhardtii
  • Application of Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating cadmium tolerance of Chlamydomonas reinhardtii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Screening of Chlamydomonas reinhardtii heavy metal cadmium-resistant mutant algal beads

[0036] (1) Preparation for inserting the target fragment

[0037]First extract the PJMG-aphⅧ plasmid (Zhangfeng Hu, Yinwen Liang, Wei He, Junmin Pan * , Cilia Disassembly with Two Distinct Phases of Regulation, Cell Reports, 2015, 10(11): 1803-1810; containing aphⅧ fragment, the sequence is shown in SEQ ID NO: 3), and then digested with EcoRI and cutsmartBuffer, and digested The product is electrophoresed on agarose gel, and the target fragment is cut and recovered, such as figure 1 As shown, M is BM5000 DNA Marker, 1 is the enzyme digestion product, and the band indicated by the arrow is the target fragment to be recovered. Gel recovery mainly uses Sangon’s Gel Recovery Kit (article number: B518191-0100). According to the weight of the gel strip, add Buffer B2 3-6 times the weight of the gel strip, and use 20 μL of Elution Buffer to elute the DNA fragments. Around 10...

Embodiment 2

[0044] Example 2: Identification of Mutant Genes of Chlamydomonas reinhardtii Heavy Metal Cadmium-Resistant Mutant Algal Strains

[0045] (1) Extraction of mutant genomic DNA

[0046] a: Put the heavy metal cadmium-resistant mutant HM24 screened above into the liquid medium with a pipette tip. After the cells grow to the logarithmic phase, collect the cells into a 1.5mL EP tube, centrifuge at 14,000rpm at room temperature for 1min, and discard. The precipitate was frozen with liquid nitrogen and stored at -80°C for future use.

[0047] b: Take out the cells collected above with liquid nitrogen, add 700 μl of preheated CTAB lysate, put them in a water bath at 65°C for about one hour, and invert and mix them several times to fully lyse the cells.

[0048] c: After the cells are fully lysed, add 700 μl of a mixed solution of phenol: chloroform: isoamyl alcohol = 25:24:1 to the EP tube, shake the EP tube, mix well, and centrifuge at 14,000 rpm for 10 min.

[0049] d: After centr...

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 an application of Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating cadmium tolerance of Chlamydomonas reinhardtii. Through inserting aph VIII segment containinga paromomycin resistance gene in Chlamydomonas at random, the Chlamydomonas reinhardtii mutated strain capable of resisting cadmium chloride obviously is acquired, the further identification finds that the cadmium resistance of the strain is caused by inserting the aph VIII segment (SEQ ID NO: 3) in Cre01.g046237.t1.1 gene. Through reducing the expression of the Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene (the CDS sequence is shown as SEQ ID NO: 1), the deactivated or deficient gene can improve the cadmium tolerance of Chlamydomonas reinhardtii; the Chlamydomonas reinhardtii strain withenhanced cadmium tolerance acquired through gene engineering method can be applied to the monitoring and governing of a cadmium polluted water area, and provides a new direction for sewage governing.

Description

technical field [0001] The invention relates to the fields of biotechnology and environmental pollution control, in particular to the application of the Chlamydomonas reinhardtii Cre01.g046237.t1.1 gene in regulating the cadmium tolerance of the Chlamydomonas reinhardtii. Background technique [0002] With the continuous improvement of modern industrialization, environmental pollution has become a global problem. Due to the unreasonable development of some factories and unreasonable industrial development, a series of environmental pollution problems have been caused, among which the most serious pollution is water and the atmosphere. The pollution of heavy metal cadmium is one of the main industrial pollution. Both industrial waste gas and waste water and automobile exhaust contain a large amount of cadmium. The nature of heavy metal pollution in the water, soil, and atmosphere will continue to accumulate, and finally the ecological environment will be destroyed, and the su...

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
IPC IPC(8): C12N15/31C07K14/405C12N1/13C02F3/32C12R1/89C02F101/20
CPCC02F3/322C02F2101/20C07K14/405
Inventor 李丽丽田广梅章伟雄田诚周俊飞胡潇曾诗乐陈斌丁梅胡长峰
Owner JIANGHAN UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap