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A kind of protein that promotes the synthesis of cotton fiber green pigment, gene and its coding sequence and application

A cotton fiber and protein technology, applied in the field of genetic engineering and genetic engineering, can solve the problems affecting the excellent properties of new materials, achieve the effect of improving the synthesis level of fiber green pigments and avoiding the introduction of non-target properties

Inactive Publication Date: 2020-09-15
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, when the traditional hybridization method transfers the green fiber gene, it will inevitably introduce some non-target traits into the new material, which will affect other excellent traits of the new material

Method used

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  • A kind of protein that promotes the synthesis of cotton fiber green pigment, gene and its coding sequence and application
  • A kind of protein that promotes the synthesis of cotton fiber green pigment, gene and its coding sequence and application
  • A kind of protein that promotes the synthesis of cotton fiber green pigment, gene and its coding sequence and application

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preparation example Construction

[0026] In the present invention, the preparation method of the coding sequence preferably includes the following steps: using the green fiber total RNA 20 days after flowering as a template, amplifying the coding sequence using a 3'-RACE kit (TaKaRa, Dalian, China). In the present invention, the sequence of the specific primer for 3'-RACE amplification is shown in SEQ ID No.32.

[0027] The present invention also provides the PCR amplification primer pair of above-mentioned coding gene or coding sequence, the nucleotide sequence of the forward primer of described PCR amplification primer pair is as shown in SEQ ID No.4, the core of reverse primer The nucleotide sequence is shown in SEQ ID No.5. The above-mentioned coding gene or coding sequence can be amplified by using the aforementioned pair of amplification primers for PCR, and the coding sequence amplified by the 3'-RACE method can be further verified.

[0028] The present invention provides a recombinant vector containin...

Embodiment 1

[0051] Fine mapping of green fiber genes

[0052] In the early stage, the green fiber (Lg) gene was assigned to the 21st chromosome (D11) with the markers C2-0120 and PGML4063 using the recombinant inbred line (RIL) population derived from the green fiber material T586 and the white fiber material Yumian 1 , HAU3033, CGR5015 and NBRI0087 are closely linked. Then, a segregation population of the F2 generation containing 750 individuals was constructed using the green fiber material (RIL051) and the white fiber variety Yumian No. This segment found 5 additional SSR markers (SWU07324, SWU01713, SWU01715, SWU16607 and SWU01720). Table 1 between molecular markers and their amplification primers used for fine mapping of green fiber (Lg) gene.

[0053] Table 1 Molecular markers used for fine mapping of green fiber (Lg) gene

[0054]

[0055]

[0056] After the cotton was mature, the fiber color of each individual plant was detected in the field, the leaves were taken to extr...

Embodiment 2

[0058] Cloning of the Gh3622 coding sequence

[0059] The total RNA of green fibers was extracted 20 days after flowering, and the cDNA sequence of Gh3622 was amplified by 3'-RACE method. The gene-specific primer for 3'-RACE was ctcttctcccataatccaattc-3', and the rest of the operations were performed according to the kit instructions (TaKaRa). The amplified products were recovered, cloned and sequenced by conventional methods.

[0060] The genome and cDNA coding sequence of the Gh3622 gene were amplified with primers 5'-ggatcc gggatgaagggcaacgat-3' and 5'-ggtacc ctaggagataggccatgac-3' using the upland cotton T586 genomic DNA and the green fiber cDNA 20 days after flowering as templates, respectively, to amplify The product was recovered, cloned, and sequenced by conventional methods, and the RACE results were further verified. The coding sequence of the finally obtained Gh3622 gene is shown in SEQ ID No.3.

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Abstract

The invention provides a protein for promoting the synthesis of cotton fiber green pigments as well as a gene, coding sequence and application of the protein, belonging to the technical fields of geneengineering and genetic engineering. The amino acid sequence of the protein provided by the invention is represented by SEQ ID No.1. By transfecting cotton through the coding gene or the coding sequence, the synthesizing level of fiber green pigments of the transfected cottons can be remarkably increased, and meanwhile, the non-target character introgression caused by virtue of a conventional hybridization measure can be avoided.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and genetic engineering, and in particular relates to a protein, a gene and its coding sequence and application for promoting the synthesis of cotton fiber green pigment. Background technique [0002] Compared with ordinary cotton, natural colored cotton has the characteristics of natural and soft color, simple and elegant, and soft texture. Textiles made of colored cotton do not need to be dyed with chemical dyes, and there will be no pollution to land and water during the processing and production process. Textiles do not contain harmful substances such as formaldehyde and azo dyes, and have anti-static and anti-itch functions. They are veritable "green products" and are known as "ecological clothing" and "plant cashmere". The new textile raw materials required by the people to protect the living environment and achieve sustainable development conform to the trend of the times when ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/74C12N1/21C12N15/82A01H5/00A01H6/60
CPCC07K14/415C12N15/74C12N15/825
Inventor 肖月华张正圣张悦严倩刘德新罗明滕中华
Owner SOUTHWEST UNIV
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