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Application of silkworm Period gene or protein in silkworm breeding and silkworm cocoon production

A silkworm and gene technology, applied in the field of sericulture, can solve problems such as tissue development without BmPeriod gene regulation

Pending Publication Date: 2022-05-06
NANTONG TEXTILE & SILK IND TECH RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the reported functions of the silkworm period gene (BmPeriod) focus on regulating circadian rhythm behavior and physiology (Ikeda et al., 2019; Sandrelli et al., 2007), and there is no report that the BmPeriod gene regulates tissue development

Method used

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  • Application of silkworm Period gene or protein in silkworm breeding and silkworm cocoon production
  • Application of silkworm Period gene or protein in silkworm breeding and silkworm cocoon production
  • Application of silkworm Period gene or protein in silkworm breeding and silkworm cocoon production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]The CRISPR / Cas9 knockout vector construction of embodiment 1 silkworm BmPeriod gene

[0035] (1) First of all, the present invention is aimed at the sequence of the BmPeriod gene, and through a large number of selections and tests, the CRISPRdirect website (http: / / crispr.dbcls.jp / ) is used to select 2 silkworms in the sixth exon of the BmPeriod gene BmPeriod gene-specific sgRNA recognition target (SEQ ID NO.3 and SEQ ID NO.4) and its full-length sequence (SEQ ID NO.5 and SEQ ID NO.6). The nucleotide sequence of the BmPeriod gene is shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2. Schematic diagram of silkworm BmPeriod gene structure and sgRNA target sequence figure 1 shown.

[0036] The sgRNA recognition target sequence is as follows:

[0037] sgRNA1: 5'-TCTCCAGCGCATGTACAGGT-3' (SEQ ID NO.3)

[0038] sgRNA2: 5'-CACTGAAGGCGACAAGGTAG-3' (SEQ ID NO.4)

[0039] The full-length sequence of sgRNA is as follows:

[0040] sgRNA1:

[0041] 5'-GACC...

Embodiment 2

[0061] Embodiment 2 Establishment of BmPeriod Gene Knockout Homozygous Line of Bombyx mori

[0062] (1) Bombyx mori egg preparation: The non-diapause eggs of silkworm Dazao were obtained by the conventional low-temperature and dark method of non-diapause egg production. Use plastic wrap as the egg attachment material, collect the eggs laid by the female moth within 30 minutes, soak in clean water for 20 minutes, and then gently wash off the silkworm eggs. Collect silkworm eggs quickly, arrange the plump and wet silkworm eggs neatly on the sterilized glass slide, absorb the water stains with filter paper, dry them for 20 minutes, put them in 3% formaldehyde water emulsion for 2 minutes, and sterilize them with water. After cleaning, let it dry for more than 20 minutes for microinjection.

[0063] (2) Introduction of the recombinant vector plasmid into silkworm eggs: the recombinant vector plasmid constructed in Example 1 was introduced into non-diapause silkworm eggs at the cl...

Embodiment 3

[0075] The silkworm BmPeriod gene mutant hybrid variety that embodiment 3 selection silk cotton production uses

[0076](1) According to the method of Example 1 and Example 2, the BmPeriod gene knockout mutant of silkworm was prepared, and the female moth homozygous for the BmPeriod gene knockout mutation was used as the female parent, and the dichroic practical hybrid combination Su with yellow cocoon The spring orange male moths of Yu × Chuncheng mated to obtain silkworm eggs of the H1 generation, and the larvae hatched from the H1 generation silkworm eggs were reared with a single moth breeding method. The low density and thick cocoon layer of the cocoon layer were used as indicators, and 10% of the moth area was preferred Mate with 10% of individuals with high cocoon silk content in the selected moth area to obtain H2 generation silkworm eggs;

[0077] (2) Raise the H2 generation larvae with a single moth breeding method, sample silkworm eggs from the embryonic development...

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Abstract

The invention relates to application of a bombyx mori Period gene or protein in bombyx mori breeding and bombyx mori silkworm cocoon production, in particular to inhibition of expression of a gene shown in SEQ ID NO.1 or protein shown in SEQ ID NO.2 during bombyx mori breeding or bombyx mori silkworm cocoon production, cultivation of a special silkworm variety for silk floss production, improvement of the thickness of a silkworm cocoon layer and the total cocoon amount of the silkworm cocoon, improvement of the silk yield of the silkworm cocoon, and improvement of the yield of the silkworm cocoon. And the content of silk fibroin in the cocoon silk fibers is increased. According to the method, the development of posterior silk glands is remarkably promoted by inhibiting the expression of the silkworm Period gene, the content proportion of silk fibroin in cocoon silk fibers is remarkably increased while the yield of silkworm cocoons is increased, the silk yield of dried cocoons is effectively increased, and the cocoon forming shape of mature silkworm larvae can be stably changed.

Description

technical field [0001] The invention relates to the technical field of sericulture, in particular to the application of a silkworm Period gene or protein in silkworm breeding and silkworm cocoon production. Background technique [0002] Sericulture production, which raises silkworms (Bombyx mori) to produce silkworm cocoons, and silk weaving production, which processes silkworm cocoons, are traditional and advantageous industries in China. More than 98% of cocoon silk of silkworm is protein, including fibrous protein silk fibroin and non-fibrous protein sericin. Cocoon silk protein is synthesized, secreted and stored in the lumen of the silk gland by the specially developed silk gland tissue in the larval stage, among which sericin protein is completely synthesized by the middle silk gland cells, and most of the silk fibroin protein is synthesized by the posterior silk gland cells, A small amount is synthesized in the posterior region of the central silk gland. The silk fi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N9/22C12N15/85C12N15/12A01K67/04A01K67/033
CPCC12N15/113C12N9/22C12N15/8509C07K14/43586A01K67/04A01K67/0333C12N2310/20A01K2207/15A01K2217/075A01K2227/70A01K2267/02
Inventor 徐世清王永峰李江岚戴泰明邱剑丰崔文沼彭如及
Owner NANTONG TEXTILE & SILK IND TECH RES INST
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