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Litopenaeus vannamei heat shock protein 40 gene lvHSP40 and application thereof in inhibition of ovary development of litopenaeus vannamei

A heat shock protein and ovarian development technology, applied in the field of aquatic organisms, can solve problems such as difficulty in synchronous development of gonads, lack of ovarian development, influence on mating and reproductive efficiency, etc., and achieve the effect of great economic value and wide application prospects.

Active Publication Date: 2019-03-15
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under artificial breeding conditions, it is difficult for the parental female and male gonads of Litopenaeus vannamei to develop synchronously, which affects the mating and breeding efficiency in production
At present, the unilateral eye-stalk removal method is used in production to promote the ovarian development of female shrimp, but there is no effective method to inhibit ovarian development.

Method used

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  • Litopenaeus vannamei heat shock protein 40 gene lvHSP40 and application thereof in inhibition of ovary development of litopenaeus vannamei
  • Litopenaeus vannamei heat shock protein 40 gene lvHSP40 and application thereof in inhibition of ovary development of litopenaeus vannamei
  • Litopenaeus vannamei heat shock protein 40 gene lvHSP40 and application thereof in inhibition of ovary development of litopenaeus vannamei

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1: Cloning and sequencing of the heat shock protein 40 gene lvHSP40 of Litopenaeus vannamei

[0022] Using the hepatopancreas tissue of Litopenaeus vannamei as material, the total RNA was extracted using the Trizol (purchased from Invitrogen Company) method, the remaining genomic DNA was digested using DNase I (purchased from Invitrogen Company), and PrimeScript was used to TM II kit (purchased from Takara Company) was reverse transcribed into the first strand of cDNA. Based on a 1075-base assembly fragment annotated as HSP40 obtained from the second-generation sequencing of the Litopenaeus vannamei hepatopancreas transcriptome (provided by Huada Genomics) on the Illumina Hiseq platform we constructed, the design gene-specific (provided by Shanghai Shenggong The company synthesized) primers were amplified to obtain the cDNA middle fragment of the heat shock protein 40 gene lvHSP40 of Litopenaeus vannamei. On the basis of obtaining the intermediate fragment, th...

Embodiment 2

[0036] Example 2: Injection of siRNA HSP40-474 inhibits gonad development of female Litopenaeus vannamei

[0037] The siRNA sequences HSP40-42, HSP40-474, and HSP40-825 were designed according to the cDNA sequence of the heat shock protein 40 gene lvHSP40 of Litopenaeus vannamei (as shown in Table 1). The siRNA was synthesized by Shanghai Sangong Company. The mRNA expression of lvHSP40 was knocked down by RNA interference by injection method, and the mRNA expression level of lvHSP40 in the hepatopancreas (Hp) and ovary (Ov) of Litopenaeus vannamei was detected at 12 hours and 24 hours after injection, and it was found that 300ng / g body weight The dose of siRNA HSP40-474 injection had the best knockdown effect on the mRNA expression of Litopenaeus vannamei lvHSP40 at each time point (as shown in Table 1). For siRNA HSP40-474, the nucleotide sequence of its sense strand is shown in SEQ ID NO.3, and the nucleotide sequence of its antisense strand is shown in SEQ ID NO.4.

[0038...

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Abstract

The invention discloses a litopenaeus vannamei heat shock protein 40 gene lvHSP40 and the application thereof in inhibition of ovary development of litopenaeus vannamei. The nucleotide sequence of thelitopenaeus vannamei heat shock protein 40 gene lvHSP40 disclosed by the invention is shown as bases from the site 174 to the site 1364 in SEQ ID NO. 1. The litopenaeus vannamei heat shock protein 40gene lvHSP40 is cloned from litopenaeus vannamei first, and experiments prove that the injection of siRNA designed by aiming at the gene is capable of effectively inhibiting the ovary development oflitopenaeus vannamei. Aiming at the problem that a method for effectively inhibiting the ovary development of litopenaeus vannamei is absent, the invention provides an effective technical means, and the litopenaeus vannamei heat shock protein 40 gene lvHSP40 has wide application prospects and extremely high economic value.

Description

Technical field: [0001] The invention belongs to the technical field of aquatic organisms, and in particular relates to the heat shock protein 40 gene lvHSP40 of Litopenaeus vannamei and its application in inhibiting the development of prawn ovaries. Background technique: [0002] Litopenaeus vannamei, commonly known as Penaeus vannamei, is native to the east coast of the Pacific Ocean. Due to its fast growth rate, strong resistance to disease and stress, low feed requirements, delicious meat, high meat yield and long-term survival time out of water Long and other advantages, it is a world-recognized excellent variety of cultured prawns. Since it was introduced from Hawaii, USA in 1988, Litopenaeus vannamei has developed into the most important species of prawn culture in my country, and its aquaculture output exceeds 70% of the total output of prawn aquaculture in China. However, under artificial breeding conditions, it is difficult for the parental female and male gonads ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113C07K14/435A61K31/713A61P15/00
CPCA61K31/713A61P15/00C07K14/43509C12N15/113C12N2310/14
Inventor 陈廷任春华黄文江晓罗鹏胡超群
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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