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A set of bac markers for distinguishing 13 pairs of chromosomes in sesame

A chromosomal and group labeling technology, applied in recombinant DNA technology, microbial assay/inspection, DNA/RNA fragments, etc., to achieve stable hybridization signals, stable and reliable hybridization results, and strong specificity.

Active Publication Date: 2021-06-04
HENAN SESAME RES CENT HENAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But in general, due to the special requirements of stability and accuracy in the identification and differentiation of sesame chromosomes, there are still many technical difficulties in the use of BAC-FISH technology for related research

Method used

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  • A set of bac markers for distinguishing 13 pairs of chromosomes in sesame
  • A set of bac markers for distinguishing 13 pairs of chromosomes in sesame
  • A set of bac markers for distinguishing 13 pairs of chromosomes in sesame

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] This example mainly introduces the screening process for the 17 BAC markers used to distinguish 13 pairs of chromosomes in sesame described in this application. The specific process is introduced as follows.

[0117] (1) Preparation of sesame chromosome specimens

[0118] (A) Sesame chromosome specimen prepared by smear method

[0119] Refer to the wall-removing and hypotonic method (Chen Ruiyang et al., The method of removing the wall and hypotonicity in the preparation of plant chromosome samples and its significance in cytogenetics, 1982) and improve it to prepare sesame chromosome samples. The specific steps are as follows:

[0120] (1) Take the healthy seeds of Yuzhi No. 11, sow them in a petri dish covered with wet filter paper, and cultivate long roots in dark at 21°C;

[0121] (2) Take a 1.2-1.8cm long root tip, cut off the top part of the 4-5mm root tip, put it in a glass bottle, add 0.002M 8-hydroxyquinoline, pretreat for 1 hour at 21°C in the dark, and tran...

Embodiment 2

[0186] In order to test the accuracy and specificity of the BAC marker group finally determined in the above-mentioned Example 1, and also to illustrate the potential use of the present invention, further, still taking Yuzhi No. 11 seed as an example, the inventors selected from 14 unknown BAC clones The BAC clones located on chromosome 2 and chromosome 4 were screened out, and the relative positions of the DNA of the BAC clone to be determined on the respective chromosomes were determined. The relevant experiments are briefly introduced as follows.

[0187] (1) Prepare the sesame chromosome sample, and refer to Example 1 for the relevant preparation process.

[0188] (2) Prepare BAC probes. When extracting BAC plasmid DNA, randomly select 14 BAC clones from hundreds of Yuzhi No. 11 BAC clones that have been identified to have specific hybridization signals (see the table below for the clone number), and screen BAC clones located on chromosome 2 and chromosome 4 were obtained....

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Abstract

The invention belongs to the technical field of sesame genomes, and specifically relates to a set of BAC marker patent applications for distinguishing 13 pairs of chromosomes in sesame. The group of BAC markers includes a total of 17 BAC information. The basic information of the BAC markers is shown in the table below, and the base sequence information of the paired ends of the BAC is shown in SEQ ID NO.1~34. The 17 BAC markers provided by this application are all single specific sites on the chromosome, with site specificity and uniqueness; using the differentiation method provided by this application, each chromosome of sesame can be accurately identified and determined, which can be used for sesame high The research on the construction of density cytogenetic map and proofreading of sesame genome has important theoretical significance and scientific value, which can provide theoretical support and lay an application foundation for subsequent sesame genomics research and sesame breeding.

Description

technical field [0001] The invention belongs to the technical field of sesame genomes, and specifically relates to a set of BAC marker patent applications for distinguishing 13 pairs of chromosomes in sesame. Background technique [0002] Sesame( Sesamum indicum L., 2n=26), is the only cultivated species in the genus Linaceae, and it is also an important characteristic high-quality oil crop in my country. The oil content in sesame seeds is 50-60%, and the ratio of oleic acid to linoleic acid is about 1:1, so it is considered to be one of the most ideal edible oils for human beings. In addition, sesame seeds also contain natural antioxidant substances such as sesamol, sesamin and vitamin E, which have important effects in health care such as lowering blood fat, antibacterial and anti-cancer, and delaying human aging. [0003] Sesame is a small-chromosomal crop with a large number of chromosomes and similar shapes, which limits the development of sesame cytogenetics researc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6841C12N15/11
Inventor 苗红梅张海洋赵瑞红马琴李春魏利斌段迎辉徐芳芳琚铭常淑娴
Owner HENAN SESAME RES CENT HENAN ACADEMY OF AGRI SCI
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