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Protein, coding genes and application thereof

A technology for encoding genes and genes, which is applied in the field of proteins and their encoding genes and applications, and can solve the problems of weakened plasma membrane selective permeability, plant cell destruction, and loss of function

Active Publication Date: 2013-04-17
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Du Tianqing et al. used Cd, Pb, and Cr to treat wheat seedlings, and found similar results. This shows that Cd, Pb, and Cr have caused serious damage to plant cells, resulting in weakened selective permeability of the plasma membrane, structural damage, and functional damage. lost

Method used

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  • Protein, coding genes and application thereof
  • Protein, coding genes and application thereof
  • Protein, coding genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the acquisition of KcPCS1 gene

[0056] 1. Acquisition of KcPCS1

[0057] 1. Total RNA extraction from candelabra

[0058] a. The mortar, spoon, tweezers, and reagent bottles used for RNA extraction were all sterilized by dry heat at 240°C for more than 2 hours; Rase-free plastic tubes and tips were purchased from Invitrogen.

[0059] b. Take 0.5ml extraction buffer (100mmol L-1 Tris-HCl, pH 8.5; 500mmol L-1EDTA.500mmol L-1 NaCl; 20% SDS; 5mmol L-1 DTT, pH=8.5; 1.5 % (v / v) β-mercaptoethanol) preheated at 65°C in a 1.5ml centrifuge tube;

[0060] c. Take 0.2g young leaves of Kadelia candel (Kc), place them in liquid nitrogen, add an appropriate amount of PVP, grind them to powder, quickly transfer them to the extract, shake the centrifuge tube vigorously, and mix them evenly;

[0061] d. Place the centrifuge tube flat at room temperature for 5 minutes, centrifuge at 12,000 rpm at 4°C for 10 minutes, and carefully transfer the supernatant to a new centrif...

Embodiment 2

[0146] Example 2, detection of heavy metal resistance of transgenic yeast

[0147] 1. Obtaining of transgenic yeast

[0148] 1. Construction of yeast expression vector

[0149] The gene sequence (sequence 1) of artificially synthesized KcPCS1 is used as a template, and PCR amplification is carried out with the following primers:

[0150] YPCS-BamHI: 5'-AGGGGATCCATGGCGGTGCCGGGCATATACA-3' (the underlined sequence is the BamHI site, sequence 3)

[0151] YPCS-SalI: 5'-GGTGTCGACTGGAAGCAATAGAGGGCGAGGAG-3' (the underlined sequence is the SalI site, sequence 4)

[0152] Take 1 μg of the above PCR product and digest it with BamHI / SalI double enzymes, and connect the obtained fragment with the pFL61 vector fragment that has also been digested with BamHI / SalI double enzymes, and transform the obtained ligation product into Escherichia coli DH5α to obtain a transformant, and extract the transformant The plasmid was identified by BamHI / SalI double enzyme digestion, and a fragment of abo...

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Abstract

The invention discloses a protein, coding genes and an application thereof. The protein is artificially synthesized and is named as KcPCS1, and is the protein as the following (1) or (2): (1) the protein consisting of amino acid sequences expressed by a sequence 2 in a sequence list; (2) the protein which is derived from (1) by substituting and / or deleting and / or adding the amino acid sequences in the sequence 2 by one or more amino acid residues and is related to stress tolerance. The experiments show that KcPCS1 gene is subjected to yeast transformation; the transgenic yeast has excellent resistance to cadmium (Cd); and the content of Cd in the body is higher than that of the yeast cell transferring into a vector.

Description

technical field [0001] The invention relates to a protein, its coding gene and application. Background technique [0002] Heavy metals generally refer to specific gravity greater than 4 or 5, density greater than 4.5g / cm 3 There are about 45 kinds of metals, such as copper, lead, zinc, iron, cobalt, nickel, vanadium, niobium, tantalum, titanium, manganese, cadmium, mercury, tungsten, molybdenum, gold, silver and other metals. Although arsenic is a non-metal element, it is often classified as a heavy metal because of its high toxicity. [0003] Excessive heavy metals have a strong toxic effect on plants and can affect the growth and development of plants. Chen Qinghua et al. used heavy metal Cr 6+ Maize seedlings were treated, and it was found that the malondialdehyde (MDA) and electrical conductivity in the plant were significantly correlated with the concentration of Cd, while the chlorophyll content and superoxide dismutase (SOD) increased with the increase of treatment...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/81C12N1/19C12N15/82
Inventor 麻密何振艳陈宏林
Owner INST OF BOTANY CHINESE ACAD OF SCI
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