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Iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as encoding gene and application thereof

A technology for transporting proteins and heavy metals, which is applied in the fields of molecular biology and genetic engineering, can solve problems that have not been reported, improve the transport capacity, improve the repair efficiency and practicability, and enhance the tolerance and enrichment of tobacco to cadmium effect of ability

Active Publication Date: 2017-08-08
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the current information about IlHMA2 in Cd 2+ The research on the mechanism of action in long-distance transport and detoxification has not been reported yet

Method used

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  • Iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as encoding gene and application thereof
  • Iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as encoding gene and application thereof
  • Iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The cloning of embodiment 1 Spine heavy metal ATPase transporter gene IlHMA2

[0028] Cultivate the water cultured to 8 weeks of horseshoe seedlings in 100mg / L CdCl 2 2.5H 2 Stress treatment in the Hoagland nutrient solution of O for 24 hours, cut the roots and wash them with sterile water, put them on sterile filter paper to absorb the water, weigh 100 mg of fresh roots, grind them fully in liquid nitrogen, and follow the TaKaRa MiniBEST Plant RNA Extraction Kit The total RNA was extracted according to the kit instructions, and the concentration was measured by a Quawell5000 nucleic acid protein analyzer, and the integrity of the RNA was detected by 1.2% formaldehyde denaturing gel electrophoresis. Take 1 μg of total RNA, and perform reverse transcription according to the instructions of the TaKaRaPrimeScriptRTase cDNA first-strand synthesis kit. , to synthesize cDNA.

[0029] According to the HMA nucleotide sequence of the plant heavy metal ATPase transporter gene pub...

Embodiment 2

[0043] Example 2 The predictive analysis of the structural characteristics of the heavy metal ATPase transporter IlHMA2

[0044] Using the TMpred program (http: / / www.ch.embnet.org / software / TMPRED_form.html) to predict the transmembrane region of IlHMA2 in S. chinensis, IlHMA2 has 8 transmembrane regions (TM1-TM8). The homology between IlHMA2 and the HMA2 proteins of wheat, rice and Arabidopsis plants was 68%, 66% and 55% respectively through the blast comparison of the NCBI website, and the function prediction of the IlHMA2 protein showed that it contained heavy metal P 1B -ATPase-specific GICCPSE, CPC, DKTGT, HP and GDGMNDAPAL motifs ( figure 1 ). Among them, the motif with metal binding domain contained in the N-terminus is GICCPSE, which belongs to the functional domain of metal-related regulation; the CPC site is responsible for metal ion conduction, and the DKTGT is the phosphorylation binding site for enzyme phosphorylation; the HP site Involved in the translocation of...

Embodiment 3

[0045] Phylogenetic tree analysis of embodiment 3 Malus sinensis heavy metal ATPase transporter gene IlHMA2

[0046] In order to analyze the phylogenetic relationship between the IlHMA2 gene and other plant HMA genes, using MEGA 6.0 software, other plants (Arabidopsis AhHMA2, Arabidopsis lyre AlHMA2, Arabidopsis AtHMA2 / 4, mustard BjHMA4, turnip BrHMA2, Rapeseed BnHMA2 / 4, Camelina CasHMA2 / 4, Clementine CcHMA2, Orange CisHMA2, Shepherd’s Purse CrHMA2, Cucumber CsHMA2 / 4, Behenia EsHMA2, Barley HvHMA2, IlHMA2 IlHMA2, Medicago truncatula MtHMA2, Celery NcHMA4 Phylogenetic tree analysis of the HMA genes of Oryza sativa, short-anther wild rice ObHMA2, rice OsHMA2, Populus euphratica PeHMA2, castor bean RcHMA2, sorghum SbHMA2, potato StHMA2, wheat TaHMA2, cacao TcHMA3, grape VvHMA3, and maize ZmHMA2). The results showed that IlHMA2 was closely related to monocotyledonous plants such as wheat TaHMA2 gene, rice OsHMA2 gene, and barley HvHMA2 gene, while its genetic relationship with dic...

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Abstract

The invention discloses an iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as an encoding gene and application thereof. Cloning is carried out for the first time to obtain the encoding gene (SEQ ID No: 1) of the iris lacteal heavy metal ATP enzyme transport protein I1HMA2, the gene I1HMA2 can be used for strengthening the tolerance or the enrichment capacity of plants to cadmium or improving the transport capacity of the cadmium in the plants, and thus the remediation efficiency of the plants to soil cadmium pollution is improved. Transgenic plants with the cadmium resistance and strong enrichment capacity are prepared by utilizing a gene engineering technology to be capable of growing in the cadmium polluted environment, so that the iris lacteal heavy metal ATP enzyme transport protein I1HMA2 as well as the encoding gene and the application thereof provide a new thought of cultivating efficient cadmium enrichment transgenic plants, have an important application prospect in the aspects of improving the soil cadmium pollution remediation efficiency and the ecological environment and provide a new way and technological support for remediation of heavy metal polluted soil.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic engineering, in particular to IlHMA2, a heavy metal ATPase transporter protein of the Sinus cuspidatum and its coding gene and application. Background technique [0002] With the rapid development of industry and agriculture in our country, the problem of soil heavy metal pollution has become increasingly serious. Among them, compared with other heavy metal elements, cadmium (Cd) is more soluble in water and has strong biological toxicity (Guo et al.2013). The reason is that Cd in the soil 2+ Easily absorbed by plant roots, resulting in a large amount of Cd 2+ Accumulated in plants, resulting in a decline in yield and quality, and more seriously, enter the human body through the food chain, posing a serious threat to human health (Clemens, 2006; Ueno et al.2010; Takahashi et al.2012). Therefore, how to effectively control and mitigate the pollution and harm of cadmium to ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8271
Inventor 郭强孟林毛培春田小霞
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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