Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment

A technology of silk protein peptide and new variety, which is applied in the field of silk protein peptide segment prepared from silkworm variety identification antibody and its preparation field

Inactive Publication Date: 2014-12-24
HUNAN NANXUN HENGYUE AGRI SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Research and develop natural colored silkworm varieties based on white silk silkworms, tussah silkworms, celestial silkworms and other natural colored silkworms, which are easy to raise and stable in inheritance. Proteins with hybrid vigor to secreted silk, but currently no method to identify them at the molecular level

Method used

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  • Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment
  • Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment
  • Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment

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Embodiment

[0049] The cloning, expression vector construction and expression of the silk non-crystalline region gene of the silkworm variety comprise the following steps:

[0050] A, according to the amino acid sequence of the non-crystalline region of the heavy chain core region of the main component of silk fibroin ( figure 1 ) and its encoding gene information, designed and synthesized (Invitrogen Company) the encoding gene sequence of the non-repeating sequence peptide SGFGPYVANGGYSGYEYAWSSESDFAG (F(1)). The 5' end and 3' end of the gene sequence contain the sticky end bases of HindIII and BglII respectively, and the recognition site of the restriction endonuclease NgoMIV is designed immediately downstream of the 5' sticky end, and immediately upstream of the 3' sticky end The recognition site of restriction endonuclease AgeI was designed, and the latter two restriction sites were used for recombination with the crystallization region gene. Select the basic plasmid containing common...

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Abstract

The invention discloses a silk protein peptide fragment for identifying a novel natural color silkworm. A gene sequence of the silk protein peptide fragment comprises domestic silkworm silk H chain amorphous region genes, recombination genes of an H chain crystalline region and an H chain amorphous region, tussah H chain genes and natural silkworm silk H chain genes. The invention further discloses a preparation method of the silk protein peptide fragment. The preparation method comprises the following steps of cloning of the silkworm silk amorphous region genes of the domestic silkworm variety and the establishment and expression of an expression vector; cloning of the recombination genes of the silkworm silk crystalline region and the silkworm silk amorphous region of the domestic silkworm variety and the establishment and expression of an expression vector; and cloning of the silkworm silk genes of the natural color cocoon variety and the establishment and expression of an expression vector. The method is simple and easy to implement, and ensures that the sequence is accurate and errorless, and the gene mutation or the gene deletion of each established expression vector is avoided, so that the triple-coded mispairing or the exceptional amino acid errors of expression products cannot be generated, and thus an obtained product can be further prepared into a silkworm silk antibody with high precision. At the present, the report related to the preparation of the peptide fragment from the white domestic silkworm silk variety (silk protein amorphous regions and crystalline regions/ amorphous region units) and the yellow tussah silk variety or green natural silkworm silk variety is not provided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a silk protein peptide (antigen) used for preparing silkworm species identification antibodies and a preparation method thereof. Background technique [0002] Silk fibroin (hereinafter referred to as silk fibroin) protein belongs to natural animal protein fiber and is a kind of structural protein, which is composed of two parts: crystalline state and amorphous state. Silk fibroin includes domestic silk and wild silk (tussah silk, celestial silk, etc.). Bombyx mori silk fibroin is a complex composed of heavy chain (H chain) protein, silk fibroin light chain (L chain) protein and glycoprotein P25. Tussah silk and celestial silk fibroin are composed of H chain. [0003] Bombyx silkworm silk fibroin has been a research hotspot for decades because of its rich sources. Scholars have conducted a lot of research on its structure, performance and application of biological materials. However,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C07K19/00C07K1/22
Inventor 王建南裔洪根褚永兴
Owner HUNAN NANXUN HENGYUE AGRI SCI & TECH CO LTD
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