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Tobacco glandular hair cDNA (complementary Deoxyribonucleic Acid) microarray for expression profile and preparation and application thereof

An expression profiling and glandular hair technology, applied in the field of gene chips, can solve the problems of inapplicability to a large number of sample studies and high sequencing costs, and achieve the effect of improving the possibility of low-abundance genes, strong specificity, and guaranteed specificity.

Inactive Publication Date: 2011-08-17
HENAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

The main disadvantage of this method is that different cDNA libraries have to be sequenced separately, so the sequencing cost is high, and it is not suitable for the study of a large number of samples

Method used

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  • Tobacco glandular hair cDNA (complementary Deoxyribonucleic Acid) microarray for expression profile and preparation and application thereof
  • Tobacco glandular hair cDNA (complementary Deoxyribonucleic Acid) microarray for expression profile and preparation and application thereof
  • Tobacco glandular hair cDNA (complementary Deoxyribonucleic Acid) microarray for expression profile and preparation and application thereof

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Embodiment 1

[0032] Example 1 A tobacco glandular trichome cDNA expression profile chip, including a slide, coated with a positively charged membrane on its plane, a dot matrix is ​​arranged on the surface of the membrane, and sample points are arranged on the dot matrix. Arranged on the array are PCR products containing 2830 cDNA clones derived from tobacco glandular trichomes, the length of which is 400-1000 bp, and contains positive controls and negative controls; the sample points are arranged in a matrix, and a single matrix is ​​arranged in 76×114 Sample points, each sample point set 3 times in the array of replicates. The positive control is the tobacco actin gene, and the negative control is the human gene that has no homology with tobacco and the blank sample dilution.

[0033] The preparation method of the above-mentioned tobacco glandular trichome cDNA expression spectrum chip specifically comprises the following steps:

[0034] (1) Collection of tobacco glandular trichomes and...

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Abstract

The invention relates to a tobacco glandular hair cDNA (complementary Deoxyribonucleic Acid) microarray for an expression profile and preparation and application thereof. A preparation method of the microarray comprises the following steps of: separating tobacco leaf glandular hairs, extracting total RNA (Ribonucleic Acid) and constructing a tobacco glandular hair cDNA library; randomly sequencing the cDNA library to obtain a tobacco glandular hair EST (Expressed Sequence Tag) database and amplifying an EST sequence by undergoing a PCR (Polymerase Chain Reaction) to obtain the tobacco glandular hair cDNA microarray for the expression profile; and analyzing tobacco glandular hairs and glandular hair removed leaf gene expression by using the microarray to obtain up-regulated expression ESTs in glandular hairs and screen a specific expression gene. The microarray obtained by the invention has the advantages of high efficiency, sensitivity, high flux, high accuracy and high specificity, and is suitable for researching tobacco glandular hairs, cultivating measures, environmental stress and the like. By adopting the microarray, the expression modes of a large number of genes can be detected at one time, and the gene expression change rules of a plurality of samples are comprehensively analyzed under the same testing condition.

Description

technical field [0001] The invention relates to the field of gene chips, in particular to a tobacco glandular trichome cDNA expression spectrum chip and its preparation and application. Background technique [0002] As a specialized structure of plant epidermal cells, plant glandular trichomes have attracted much attention because of their important roles in plant pest defense, adversity stress resistance and secondary product synthesis. With the advancement of functional genomics technology, the use of microarray technology to study the molecular mechanism of plant glandular hair development, metabolism and secretion has gradually become a hot spot in the field of plant biology research. Due to the complete genome information and perfect microarray products of Arabidopsis thaliana, there are many favorable conditions for the molecular biology research of its glandular trichomes. However, since Arabidopsis is a typical non-secreting glandular trichome, these favorable condi...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 崔红张松涛杨慧娟叶协锋冀浩靳超肖炳光刘国顺
Owner HENAN AGRICULTURAL UNIVERSITY
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