STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of STR molecular markers

A technology of molecular markers and sporotrichum, applied in DNA/RNA fragments, biochemical equipment and methods, microorganisms, etc., can solve the problems of few applications, few microsatellite sites, and difficulty in microsatellite site information

Active Publication Date: 2017-11-24
ICDC CHINA CDC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In terms of pathogenic fungi, there are relatively few applications at present. One of the important reasons is that

Method used

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  • STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of STR molecular markers
  • STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of STR molecular markers
  • STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of STR molecular markers

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Screening / acquisition and primer design of Sporothrix globosa STR loci

[0030] Based on the genome sequencing results of Sporotrichum globosa (GenBank: LVYW00000000.1), 200 microsatellite loci were isolated from the genome sequencing data using SciRoKo software. After screening, a total of 150 pairs of microsatellite-related primers were designed by Primer premier 5.0 software, and the related primers were initially verified by e-PCR. A strain of Sporotrichum globosa was selected, and the above primers were verified by gradient PCR (while determining their appropriate annealing temperature), and 107 pairs of primers were able to amplify stably. Ten strains of Sporotrichum globosa from different sources were selected, and after genetic polymorphism detection, 25 STR loci with stable amplification and clear bands were finally obtained, as shown in Table 1.

Embodiment 2

[0031] The population genetic diversity detection of embodiment 2 Sporotrichum globosa

[0032] 1. Genomic DNA extraction of Sporotrichum globosa

[0033] The yeast genome DNA extraction kit (CW0569S) purchased from Beijing Kangwei Century Biotechnology Co., Ltd. was used to complete the genome DNA extraction of 22 strains of Sporothrix globosa, and the operation procedure was carried out according to the kit instructions. 22 strains of Sporotrichum globosa were provided by the Mycology and Mycosis Research Center of Peking University.

[0034] 2. Microsatellite PCR amplification

[0035] The microsatellite site-specific primers of the present invention are used to amplify the 22 samples of Sporothrix globosa extracted in the step. The microsatellite marker-specific primer of Sporothrix globosa was fluorescently labeled by FAM labeling the 5' end of the forward primer with a fluorescent dye.

[0036] The PCR reaction system is 20 ul, including 10 ul of 2×Premix Taq (TAKARA,...

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Abstract

The invention relates to STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of the STR molecular markers. The invention provides a microsatellite locus (STR molecular marker) combination for the Sporothrix globosa for the first time and provides a set of specific primers (SEQ ID NO:26-75) capable of efficiently amplifying Sporothrix globosa populations as well as detection conditions. The microsatellite locus combination contains 25 highly polymorphic microsatellite loci (SEQ ID NO:1-25). The set of microsatellite loci for the Sporothrix globosa not only can be applied to genetic background analysis and molecular tracing of the Sporothrix globosa, but also can be applied to molecular epidemiology, population genetics and phylogenetic analysis of the Sporothrix globosa.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to the STR molecular marker of Sporothrix globosa and its application. Background technique [0002] Sporotrichosis is a subacute or chronic fungal infection of the subcutaneous tissue, mainly manifested as chronic granulomatous lesions of the skin, subcutaneous tissue, and nearby lymphatic system, which can spread through the blood and lymphatic system and involve the heart, lungs, brain, etc. Organs have a certain rate of disability and death. In my country, sporotrichosis is a deep fungal disease second only to candidiasis and aspergillosis. It is mainly prevalent in the three eastern provinces, but it is distributed throughout the country. [0003] The main pathogenic fungi that cause sporotrichosis include: Sporothrix schenckii, Sporothrix globosa, Sporothrix brasiliensis, Sporothrix mexicana, Leucospore Sporothrix albicans and Sporothrix schenckii var. lu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 龚杰李若瑜张建中何利华万喆
Owner ICDC CHINA CDC
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