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cDNA chip for determining gene expression information of crucifer under threatening conduction

A gene expression and cruciferous technology, applied in the field of cDNA chips, can solve the problem of large target fragments

Inactive Publication Date: 2007-04-25
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Conventional hybridization detection methods mainly include target fragment amplification, direct spotting or transfer to nylon membrane after electrophoresis, and UV hinge. has certain limitations

Method used

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  • cDNA chip for determining gene expression information of crucifer under threatening conduction

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Experimental program
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Effect test

Embodiment 1

[0022] Example 1. Preparation of a cDNA chip for detecting gene expression information of cruciferous plants under stress conditions

[0023] The preparation method of the cDNA chip comprises the following steps:

[0024] 1. Construction of suppression subtractive library and cDNA library of Chinese cabbage in rosette stage and heading stage of Chinese cabbage infected by Erwinia soft rot pathogen

[0025] Firstly, the PCR-SelectTM cDNA Subtraction kit (Clontech, CA, USA) was used to construct the suppression and subtraction library of Chinese cabbage leaves infected by Erwinia soft rot pathogen according to the kit instructions, and the SuperscriptTM Lambda System (GibcoBRL) was used to construct according to the kit instructions. The cDNA libraries of Chinese cabbage at the rosette stage and the heading stage; then clones were randomly selected from the two libraries for sequencing.

[0026] 2. Select non-redundant sequences for making probes

[0027] The EST that step 1 o...

Embodiment 2

[0034] Embodiment 2, use the cDNA chip of the present invention to detect the difference of high resistance and high sensitivity turnip mosaic virus Chinese cabbage inbred lines (both F6 generation) to turnip mosaic virus reaction

[0035] Detection of high resistance and high sensitivity turnip mosaic virus Chinese cabbage inbred lines (both F6 generation) with cDNA chip of the present invention is to the difference of turnip mosaic virus reaction, concrete experimental procedure and result are as follows:

[0036] 1. Drawing materials

[0037] When the plants of Chinese cabbage inbred lines with high resistance and high sensitivity to turnip mosaic virus (both F6 generation) grow to 4-5 true leaf stage, the leaves infected with TuMV virus stored at -20 ℃ are treated with pH7.0 , phosphate buffer solution with a concentration of 0.1M was ground at 1:5 (wt / vol), and the supernatant liquid was taken for inoculation. Turnip mosaic virus (TuMV) was inoculated on the newly develo...

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Abstract

The invention discloses a cDNA chip for detecting gene expressing information of crucifer plants under biological or non-biological stress. The gene chip takes the cDNA fragment of Chinese cabbage gene under versity and special growing stage as the probe which is composed by probe with sequences shown in SEQ ID No.:1-221DNA and probe sequence from 3885 strips gene bank. The invention can be used not only in batch-detecting of gene expressing information of crucifer plants under biological or non-biological stress, screening of differential gene, but also can be used to build gene expressing profiles of cruciferous plants in different growing stages, different tissues and organs, and even idioblast. High-throughput detection of gene expression information can be carried out by the application of the gene chip, which is convenient and safe, with reliable and effective test results, and can save a lot of time, human, material and financial resources and has broad application prospects.

Description

technical field [0001] The invention relates to a cDNA chip for detecting gene expression information of plants under stress conditions, in particular to a cDNA chip for detecting gene expression information of cruciferous plants under biological stresses such as diseases and insects and abiotic stresses such as drought and cold. Background technique [0002] Using known gene information to specifically detect the expression information of the target gene in the sample to be tested is a commonly used detection method in the field of molecular biology, such as the commonly used Southern hybridization, Northern hybridization and PCR techniques. However, with the development of molecular biology, these techniques cannot simultaneously analyze thousands of genes under the same conditions, so they have certain limitations. It has become a trend to immobilize thousands of genes on the carrier by in situ synthesis (in situ synthesis) probes or synthesis of target gene fragments and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 马荣才张松贺常凤启谢华屈淑平远方戴大鹏
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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