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AOX specific antibody combination and preparation method and application thereof

A specific and antibody technology, applied in the field of plant cells, can solve the problems of difficult level, difficult and difficult detection of AOX1a, etc., and achieve the effect of improving detection efficiency.

Active Publication Date: 2020-12-29
NORTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research focuses on AOX1a, and there are few other four reports. AOX1a is the main one, and its expression is relatively large, while the other four are difficult to detect. Generally used antibodies are from UNIVERSITY OF NEBRASKA, LINCOHN, which is A monoclonal antibody with good specificity, but the sensitivity of this antibody is very low, even AOX1a is difficult to detect
[0004] In the western blot experiment (Western Blot) to detect the expression of AOX, the membrane protein of the cell must be extracted first, and then the AOX antibody with a dilution factor of 1:50 can be used to display AOX1a by Western Blot, because the other four AOX The abundance of AOX1a is far less than that of AOX1a, and it is very difficult to detect their levels in plants, and this antibody cannot distinguish which subtype of AOX, it detects the whole of the AOX family

Method used

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  • AOX specific antibody combination and preparation method and application thereof
  • AOX specific antibody combination and preparation method and application thereof
  • AOX specific antibody combination and preparation method and application thereof

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Embodiment 1

[0028]This embodiment provides a method for preparing AOX specific antibody combination, including the following steps:

[0029]S1, synthesis of target gene sequence:

[0030]

[0031]As shown in SEQ ID NO.1;

[0032]The amino acid sequence of the antibody AOX1a is shown in SEQ ID NO. 5: ASTITLGE KTPMKEEDANQKKTENESTG GDAAGGNNKG DKGIA;

[0033]

[0034]As shown in SEQ ID NO.2;

[0035]The amino acid sequence of the antibody AOX1b / c is shown in SEQ ID NO. 6: SKMTF EKKKTSEEEEGSGDGVKVND QGNKGEQLIV;

[0036]

[0037]As shown in SEQ ID NO.3;

[0038]The amino acid sequence of the antibody AOX1d is shown in SEQ ID NO. 7: LSSDTSSPV SGNNQPENPIRTADGKVIST YWGIP;

[0039]

[0040]As shown in SEQ ID NO.4;

[0041]The amino acid sequence of the antibody AOX2 is shown in SEQ ID NO. 8: GMSSASAM EKKDENLTVKKGQNGGGSVA VPSYWGIETA;

[0042]In the target gene sequence SEQ ID NO.1-SEQ ID NO.4, the single-underlined base part is the protected base, the double-underlined base part is the EcoRI and XhoI restriction site, and the remaining base part ...

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Abstract

The invention relates to the technical field of plant cells, and particularly discloses an AOX specific antibody combination and a preparation method and application thereof. The AOX specific antibodycombination comprises four antibodies including AOX1a, AOX1b / c, AOX1d and AOX2. The obtained AOX specific antibody combination is very high in sensitivity, and the four AOX specific antibodies can respectively detect AOX proteins with highly similar protein sequences in plants.

Description

Technical field[0001]The present invention relates to the technical field of plant cells, in particular to AOX specific antibody combinations and preparation methods and uses thereof.Background technique[0002]Photosynthesis is the main source of the earth's energy. Research on photosynthesis can help increase crop yields and may also provide new solutions to the problem of energy shortages. Plant photosynthesis involves the electron transport chain, and AOX protein is the terminal oxidase of the alternative oxidase electron transport pathway. It participates in cyanide-resistant respiration and plays a very important role in the process of plant resistance to adversity stress. Therefore, it has become a research hotspot.[0003]There are five AOX proteins in plant mitochondria: AOX1a, AOX1b, AOX1c, AOX1d and AOX2. They are homologous and have a high degree of protein similarity. The current research focuses on AOX1a, and the other four reports are few. AOX1a is the main one. Relativel...

Claims

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Application Information

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IPC IPC(8): C07K16/40C07K16/06C12N15/13C12N9/02C12N15/53C12N15/70G01N33/573
CPCC07K16/40C07K16/065C12N9/0004C12N15/70G01N33/573G01N2333/902Y02A50/30
Inventor 王丹凤付爱根王春宇
Owner NORTHWEST UNIV
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