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SSR DNA markers for jute expressed sequence tags

A technology for expressing sequence tags and DNA markers, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., and can solve the problems of limited research on jute microsatellites

Inactive Publication Date: 2014-07-23
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on jute microsatellites in my country is still very limited, and only a handful of laboratories have started related work

Method used

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  • SSR DNA markers for jute expressed sequence tags
  • SSR DNA markers for jute expressed sequence tags
  • SSR DNA markers for jute expressed sequence tags

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1:Sixty-six new microsatellite markers suitable for the analysis of jute genetic diversity were developed. The 66 microsatellite marker numbers are respectively: CcSSR001-CcSSR045 (Jute EST-SSR), CoSSR046-CoSSR066 (Jute EST-SSR), as shown in Table 1.

Embodiment 2

[0026] Example 2: Ten polymorphic microsatellite markers CcSSR008, CcSSR013, CcSSR016, CcSSR024, CcSSR029, CcSSR040, CoSSR049, CoSSR050, CoSSR052 and CoSSR053 were identified. as table 3 and figure 1 shown.

[0027] Table 3 Polymorphism and amplification efficiency of jute EST-SSR

[0028]

[0029]

[0030]

[0031] ABCD indicates the level of clarity of the electrophoretic band pattern.

Embodiment 3

[0032] Example 3: Genetic diversity analysis of the genus Jute was analyzed using the microsatellite markers developed above. The test materials were 48 pieces of jute long-fruited wild species jute, long-fruited wild species jute, long-fruited cultivated species, round-fruited cultivated species jute and round-fruited wild species jute from 11 countries and regions (Table 4). . Using NTsys software to calculate the genetic similarity coefficient of 48 jute materials. The calculation method is unweighted cluster analysis (UPGMA), and the reading band adopts the method of 0 and 1, 0 means no band, and 1 means band. The results of cluster analysis showed that these 10 microsatellite markers could be well used in the genetic diversity analysis of jute. The results of cluster analysis are as follows: figure 2 As shown, 48 jute materials are divided into two categories, one is round fruit species and the other is long fruit species.

[0033] Table 4 Variety names and sourc...

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Abstract

The invention relates to a molecular marker technology, specifically to SSR DNA markers for jute expressed sequence tags. A preparation method for the markers comprises the following steps: developing novel SSR sites; amplifying primer sequences of the SSR sites; and carrying out polymorphic analysis on the SSR sites. The invention is characterized in that jute expressed sequence tags are downloaded from the GenBank common database, 66 novel SSR markers of jute are separated and identified, 10 SSR markers with rich polymorphism are selected from the 66 novel SSR markers, and the 10 SSR markers comprise CcSSR008, CcSSR013, CcSSR016, CcSSR024, CcSSR029, CcSSR040, CcSSR049, CcSSR050, CcSSR052 and CcSSR053. The invention provides 66 novel jute SSR sites and the primer sequences and an amplification method used for amplifying the 66 SSR sites, which can be used for analysis of genetic diversity of different species of jute, construction of a genetic linkage map, molecular marker-assisted breeding and other research.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a jute expression sequence tag microsatellite DNA molecular marker. Background technique [0002] Microsatellites (also known as simple repeat sequences, SSRs) are short sequences of 1-6 nucleotides that repeat multiple times in series; among them, dinucleotides and trinucleotides are the most common Repeating units such as (CT)n, (AT)n, (ACC)n, (CGA)n and (AGA)n. Because the flanking sequences of microsatellites are relatively conservative, primers can be designed according to the flanking sequences, and the sequences of microsatellite loci can be amplified by PCR. Due to the different repeating times of repeating units in microsatellites, the length of amplified microsatellite sequences is polymorphic. Microsatellite sequences widely exist in eukaryotic genomes, and are randomly distributed, with high polymorphism, good repeatability and stability. They are very effective mol...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 张立武祁建民徐建堂陶爱芬林荔辉方平平林培清吴建梅
Owner FUJIAN AGRI & FORESTRY UNIV
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