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

Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof

A gene and protein technology, applied to the gene DvCRP2 with anti-Cd2+ and anti-Cu2+ functions, its encoded protein and application fields, can solve problems such as inability to study and lack of transformation system in salina

Inactive Publication Date: 2012-03-28
SHANGHAI UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on salina lacks a transformation system and cannot be studied by mature genetic methods

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
  • Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof
  • Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof
  • Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Cloning and analysis of the full-length coding region of DvCRP2

[0041] The yeast expression library was transformed into the yeast mutant Δyap1 by LiAc heat shock method, in the presence of 150 μM Cd 2+ The yeast transformants with mutant phenotype recovery were screened under the screening pressure to obtain candidate clones. In order to exclude false positives caused by spontaneous mutations in the yeast genome, the expression vectors carried by the candidate clones were isolated from yeast cells, transformed into Escherichia coli for amplification, and the amplified vectors were reverted to the yeast mutant Δyap1 and incubated in 150 μM Cd 2+The clones that can still grow stably under the treatment are the true positive clones. The insert fragment in the expression vector carried by the positive clone was sequenced to obtain the partial cDNA sequence of DvCRP2.

[0042] In order to obtain the cDNA sequence of the full-length coding region, the present ...

Embodiment 2

[0043] Example 2 Sequence analysis of DvCRP2

[0044] The full-length cDNA sequence of the DvCRP2 gene was searched for homology in the corresponding database of NCBI using Blastx, tBlastx and Blastn programs successively ( http: / / blast.ncbi.nlm.nih.gov ), neither yielded valid Blast results ( figure 2 ), indicating that the salina DvCRP2 gene and its homologous genes have not been cloned and studied at present, and are discovered for the first time with anti-Cd 2+ Functional new genes.

Embodiment 3

[0046] Example 3 Functional Analysis of DvCRP2 in Yeast

[0047] (1) Construction of yeast expression vector

[0048] The full-length ORF fragment of DvCRP2 was amplified by using the cDNA clone of DvCRP2 as a template by PCR method. In order to construct the clone, with the help of primer introduction method, an EcoRI restriction site is added to the 5' end of the target sequence, and an XhoI restriction site is added to its 3' end. The primer sequences are as follows:

[0049] DvCRP2+: 5'-attgaattcatgatgcatgcagaccagcg-3'

[0050] DvCRP2-:5'-atctcgagtcacagtcccacatgcctct-3'

[0051] The product amplified by PCR was directional cloned into the vector pAJ401 using the enzyme sites EcoRI and XhoI, and transformed into Escherichia coli Top10. The positive clone pAJ401-DvCRP2 was identified by enzyme digestion and sequencing.

[0052] (2) Transform yeast

[0053] Using the LiAc heat shock transformation method: the empty vector pAJ401 and the expression vector pAJ401-DvCRP2 of...

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 a cDNA sequence of a Dunaliella viridis Cd2+ resistant and Cu2+ resistant gene DvCRP2 and an amino acid sequence for encoding protein, and relates to function and application of the gene. The invention adopts a yeast functional complementation system to screen to obtain the DvCRP2 gene, applies a RACE technology to obtain the overall-length cDNA sequence, and proves that the gene can greatly improve the Cd2+ resistant and Cu2+ resistant capabilities of model organism yeast and significantly increase the accumulation amount of cadmium in yeast cells. The disclosed overall-length gene and the amino acid sequence are first discovered, which are Cd2+ resistant and Cu2+ resistant genes peculiar to Dunaliella viridis. The overall-length gene has application value in improving Cd2+ resistant and Cu2+ resistant capabilities of an organism, increasing accumulation amount of cadmium and copper in the organism in gene engineering aspects.

Description

technical field [0001] The present invention relates to a kind of anti-Cd 2+ and anti-Cu 2+ Functional gene DvCRP2, its encoded protein and its application. Background technique [0002] With the development of modern industry, heavy metal pollution is becoming more and more serious. Environmental and health problems caused by heavy metal pollution have been reported in many countries. Cadmium (Cd 2+ ) is one of the most toxic heavy metals, and Cd 2+ In the environment, it has strong chemical activity, high mobility and long-lasting toxicity. Since the 1920s, due to the rapid development of mining, smelting, electroplating, chemical and other industries and the discharge of wastewater, water and soil Cd 2+ Pollution is getting worse. Contaminated Cd in water and soil 2+ It is easily absorbed and accumulated by aquatic organisms and higher plants, affecting their growth and metabolism, resulting in a decline in the yield and quality of agricultural products and damage...

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): C12N15/31C12N15/10C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C07K14/405
Inventor 宋任涛许政暟孟祥宗余浣莎
Owner SHANGHAI 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