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Method for increasing content of carotenoid in patinopecten yessoensis

A technology of carotene and scallop, which is applied in DNA/RNA fragments, animal husbandry, recombinant DNA technology, etc., and can solve the problems of decreased expression level and reduced degradation efficiency of carotenoids

Pending Publication Date: 2019-06-04
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The expression of this gene is significantly decreased, which reduces the degradation efficiency of carotenoids, which is the reason for the accumulation of carotenoids in the orange adductor muscle

Method used

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  • Method for increasing content of carotenoid in patinopecten yessoensis
  • Method for increasing content of carotenoid in patinopecten yessoensis
  • Method for increasing content of carotenoid in patinopecten yessoensis

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Experimental program
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Embodiment 1

[0016] Embodiment 1: screening protease

[0017] Through genome-wide linkage analysis, association analysis, expression profile analysis and other multi-omic studies, the applicant found that the expression of carotenoid oxidative lyase gene is down-regulated, which reduces the degradation efficiency of carotenoids. The reason for the red.

[0018] Finally, the target gene was obtained from the scallop genome and transcriptome database. The gene has a full length of 9107bp, a cDNA length of 2682bp, and a CDS length of 1566bp (SEQ ID NO: 2), encoding 521 amino acids (SEQ ID NO: 1), predicted The molecular weight of the protein is 59.2KDa, the isoelectric point is 6.54; the length of 5'UTR is 510bp, and the length of 3'UTR is 609bp.

[0019] By injecting dsRNA, the gene expression is interfered with. qRT-PCR was used to detect the effect of interference. By observing the color change of adductor muscle and HPLC analysis, it was proved that carotenoids accumulated in the muscle...

Embodiment 2

[0021] Embodiment 2: Improve the content of carotenoids in Ezo scallop by RNAi interference

[0022] The RNA interference of the gene of the present invention comprises the following steps:

[0023] a) primer design;

[0024] b) preparation of RNAi template;

[0025] c) in vitro transcription and synthesis of dsRNA;

[0026] d) Injecting dsRNA to interfere with the expression of the target gene;

[0027] e) qRT-PCR detection interference effect;

[0028] f) HPLC detection of carotenoid content.

[0029] The specific operation is as follows:

[0030] a) Primer design. The RNAi target sequences of genes were predicted and screened using siDirect version 2.0 (http: / / sidirect2.rnai.jp / ). According to the basic principles of RNAi primer design, corresponding primers were designed for the target sequence, and a T7 promoter was introduced at the 5' end of each primer. The primer sequence was:

[0031] 5'-TAATACGACTCACTATAGGGCTCCCTTCGATGTAGCTGAAAAATT-3';

[0032] 5'-TAATACGAC...

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Abstract

The invention provides a method for increasing the content of carotenoid in patinopecten yessoensis, which is used for increasing the content of carotenoid in adductor muscles of scallops by inhibiting the expression quantity of protease having the function of degrading carotenoid in the patinopecten yessoensis. The method is performed by reducing the quantity of protease having an amino acid sequence of SEQ ID NO:1 in patinopecten yessoensis; and a RNA interference technology is used for inhibiting expression of a target gene and detecting variation of the expression quantity of the gene after interference and the content of carotenoid in adductor muscles of scallops, which proves that screened protease has the function of degrading the carotenoid in patinopecten yessoensis, and the content of carotenoid in adductor muscles of scallops can be increased by inhibiting the expression of the protease.

Description

technical field [0001] The invention belongs to the technical field of functional gene screening, and in particular relates to a method for increasing the carotenoid content of scallops. Background technique [0002] Patinopecten yessoensis was introduced into my country from Japan in the 1980s and has become an important marine cultured shellfish in northern China. The adductor muscles of common Ezo scallops are white, but some individuals in the cultured populations of Ezo scallops in my country have orange-red adductor muscles, and these individuals are normal in terms of growth, development and reproduction. Studies have shown that the accumulation of two carotenoids, scallopolone and combxanthin, is the reason for the orange-red color of the scallop adductor muscle, while scallopolone and combxanthin cannot be detected in the white adductor muscle of common Ezo scallops . Further family analysis showed that the orange-red color of adductor muscle is a recessive trait ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A01K67/033
Inventor 胡晓丽包振民王姝玥赵亮李雪张梦燃王慧贞刘诗琪陆维
Owner OCEAN UNIV OF CHINA
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