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Molecular marker and method for identifying excellent stichopus japonicus selenka variety

A technology of molecular markers and excellent varieties, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., to achieve the effect of improving breeding efficiency, shortening breeding time, and rapid identification

Inactive Publication Date: 2013-02-06
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no molecular marker report for the screening of fine varieties of Apostichopus japonicus

Method used

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  • Molecular marker and method for identifying excellent stichopus japonicus selenka variety
  • Molecular marker and method for identifying excellent stichopus japonicus selenka variety
  • Molecular marker and method for identifying excellent stichopus japonicus selenka variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] In March 2009, 20 live sea cucumbers of about 3g / piece were purchased from Xiangshan Port, Ningbo City, Zhejiang Province, and kept in the laboratory for 2 weeks, changing 2 / 3 volume of seawater every day, and each sea cucumber was followed by the following method Quickly identify whether the sea cucumber variety is good or not:

[0014] 1. Take 1.0 mL of body cavity fluid of sea cucumber, centrifuge at 2000 g for 5 min, collect blood cells, add 1.0 mL of Trizol reagent (purchased from Clontech Company), shake and mix, leave at room temperature for 5 min, then add 0.2 mL of chloroform, shake and mix, and let stand at room temperature 10min, 4°C, 12000rpm, centrifuge for 15min, draw the supernatant into a PE tube, add an equal volume of isopropanol to the supernatant, mix well, let stand at room temperature for 5min, 4°C, 12000rpm, centrifuge for 10min, remove the supernatant, and Add 1 mL of ethanol with a mass percentage concentration of 75% to the precipitation, centr...

Embodiment 2

[0019] The gene fragments of the above-mentioned ADP glycosylation factors, arginine kinase, cytochrome oxidase c subunit, heat shock protein 70, and actin were sequenced separately, and the sequences were identified by Blast software to obtain the gene fragments of ADP glycosylation factors The nucleotide sequence is shown in SEQ ID NO.1, the gene fragment nucleotide sequence of arginine kinase is shown in SEQ ID NO.2, and the gene fragment nucleotide sequence of cytochrome oxidase c subunit is shown in SEQ ID Shown in NO.3, the gene fragment nucleotide sequence of heat shock protein 70 is shown in SEQ ID NO.4, and the gene fragment nucleotide sequence of actin is shown in SEQ ID NO.5, proves that the present invention designs The specificity of each primer.

Embodiment 3

[0022]

[0023]

[0024]

[0025]

[0026]

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Abstract

The invention discloses a molecular marker and method for quickly identifying the excellent stichopus japonicus selenka variety. The primers of the molecular marker are the primers of the amplified ADP glycosylation factor, amplified arginine kinase, amplified cytochrome oxidase c subunit, amplified heat shock protein 70 and amplified primer actin. The expressing genes of the molecular marker are the gene fragments of the ADP glycosylation factor, arginine kinase, cytochrome oxidase c subunit, heat shock protein 70 and actin. The method comprises the following steps: extracting the total RNA, synthesizig the cDNA, and amplifying and identifying by PCR (polymerase chain reaction). The molecular marker and the method can be used for identifying quickly, easily, specifically and efficiently. If the expression level of at least three of the gene fragments is 2.0, the stichopus japonicus selenka is the excellent variety which can grow up quickly. Therefore, the stichopus japonicus selenka which can grow up quickly can be screened out and cultured as the excellent variety, the culturing time of the stichopus japonicus selenka can be shortened, and the culturing benefit of the stichopus japonicus selenka can be improved.

Description

technical field [0001] The invention relates to biomolecular markers, in particular to molecular markers for identifying fine varieties of sea cucumbers and a rapid identification method. Background technique [0002] Sea cucumber (Stichopus japonicus Selenka) belongs to the phylum Echinodermata, the class Aposthopae, and the order Aropodactylus, commonly known as sea mice and sea cucumbers. It is mainly distributed in the coastal areas of Dalian, Beidaihe, Shandong Peninsula, Lianyungang and Pingshan Island in Jiangsu. Beginning in 2006, we introduced northern sea cucumbers into the shrimp ponds in the Xiangshan area of ​​our city for cultivation. The average weight of sea cucumbers that have been cultured for about 6 months can increase from 3g / seedling to 50-60g, and the survival rate is relatively high. In the process of sea cucumber cultivation in the south, we found that the growth speed of sea cucumber species is significantly different. There are fast-growing variet...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 李成华朱琳苏秀榕金春华李太武
Owner NINGBO UNIV
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