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A kind of tobacco nbplp gene polyclonal antibody and its preparation method and application

A polyclonal antibody, Bunsen cigarette technology, applied in biochemical equipment and methods, botanical equipment and methods, applications, etc.

Active Publication Date: 2022-04-22
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

NbPLP is a member of the chlorophyll a / b binding protein family, which is involved in the assembly and repair of the mechanism of photosynthesis during the development of early chloroplasts and abiotic stress, but there is no report on the antibody of NbPLP protein

Method used

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  • A kind of tobacco nbplp gene polyclonal antibody and its preparation method and application
  • A kind of tobacco nbplp gene polyclonal antibody and its preparation method and application
  • A kind of tobacco nbplp gene polyclonal antibody and its preparation method and application

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Embodiment Construction

[0041] The present invention will be further described below in combination with specific embodiments and accompanying drawings.

[0042] (1) Extraction of total RNA from tobacco leaves

[0043] The seeds of N. benthamiana were preserved in this laboratory, and the total RNA of N. benthamiana leaves was extracted by Trizol method, and then 2% agarose gel electrophoresis was carried out to detect the quality of the total RNA of N. benthamiana leaves extracted. figure 1As shown, it can be seen that the integrity of the total RNA extracted from tobacco leaves is particularly good, without degradation or contamination by DNA, and can be used as an ideal template for reverse transcription in the next step.

[0044] (2) reverse transcription

[0045] The total RNA of tobacco leaves obtained in the above step was used as a template for reverse transcription to obtain cDNA.

[0046] (3) Design and synthesis of NbPLP gene primers

[0047] ①Primer design

[0048] According to the s...

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Abstract

The invention discloses a tobacco NbPLP gene polyclonal antibody and its preparation method and application, comprising (1) constructing a recombinant expression vector containing the nucleotide sequence shown in SEQ ID NO.1, transforming the recombinant expression vector into Escherichia coli competent cells obtain NbPLP protein; (2) use the NbPLP protein as an antigen to immunize animals, obtain NbPLP protein polyclonal antibody through serum separation and purification, and utilize the polyclonal antibody prepared by the method provided by the present invention to specifically recognize The NbPLP of Nicotiana benthamiana has wide application in the detection, functional identification and research of the NbPLP protein of Nicotiana benthamiana. The preparation of NbPLP protein antibody is of great significance for detecting the expression of NbPLP gene in Nicotiana benthamiana during virus infection and studying the function of NbPLP protein.

Description

technical field [0001] The invention belongs to the technical field of biotechnology products for plant virus detection, and specifically relates to a polyclonal antibody of NbPLP gene of tobacco benthamiana and its preparation method and application. Background technique [0002] Native to Australia, Nicotiana benthamiana, like peppers, tomatoes, potatoes and cultivated tobacco, belongs to the family Solanaceae (Solanaceae). It is an allotetraploid plant with 19 chromosomes. There is no complete genome annotation yet data. Nicotiana benthamiana is one of the important model plants, which has been widely used in plant-microbe interaction, identification of protein interaction and subcellular localization. Therefore, the study of its functional genes is of great significance to the development of molecular biology. [0003] Nicotiana benthamiana light-induced protein, NbPLP for short. NbPLP is a member of the chlorophyll a / b-binding protein family, which participates in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/70C07K14/415C07K16/16C07K16/06G01N33/68
CPCC07K14/415C07K16/16C07K16/065C12N15/70G01N33/68G01N2333/415
Inventor 张坤庄新建陈佳欢徐红梅贺振甘海峰
Owner YANGZHOU UNIV