Identification and application of cotton fibre-specific HB (homeobox) transcription factor gene KNAT7

A gene and gene coding technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of the size of the HB gene and the difference in the structure of the encoded protein.

Inactive Publication Date: 2013-07-10
HUAZHONG NORMAL UNIV
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Problems solved by technology

A large number of HB genes found in plants are different in gene size, encoded protein structure, etc.

Method used

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  • Identification and application of cotton fibre-specific HB (homeobox) transcription factor gene KNAT7
  • Identification and application of cotton fibre-specific HB (homeobox) transcription factor gene KNAT7
  • Identification and application of cotton fibre-specific HB (homeobox) transcription factor gene KNAT7

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Embodiment Construction

[0040] 1. Isolation and cloning of full-length cDNA of cotton KNAT7 gene

[0041] A new gene of the KONX family was isolated from the cotton fiber cDNA library (according to Li XB et al., 2002, Molecular characterization of the cotton GhTUB1 gene that is preferentially expressed in fiber.Plant Physiol 130: 666-674), named as KNAT7.

[0042] 2. Cloning of the full-length DNA sequence of the KNAT7 gene

[0043] Design a pair of primers (see Table 1) at both ends of the KNAT7 gene ORF, use cotton genomic DNA as a template, at 94°C-5 minutes, 1 cycle; 94°C-1 minute, 58°C-1 minute, 72°C-2 Minutes, a DNA fragment of about 1 kb was obtained under 32 cycles of PCR reaction conditions, and the fragment was inserted into the PBS-T vector. The target DNA fragments were sequenced using the T3 and T7 promoters on the PBS plasmid. The obtained sequence was analyzed by DNAstar software, compared with the cDNA sequence, and the size and insertion position of the intron were analyzed.

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Abstract

The invention discloses the identification and application of a cotton fibre-specific HB (homeobox) transcription factor gene KNAT7. The cotton fibre-specific KNAT7 belongs to a member of the homeobox (HB) transcription factor gene family, the gene sequence does not have an intron, and the overall length of the cDNA (complementary DNA) of the cotton fibre-specific KNAT7 is 1244bp. A KNAT7 gene-coded protein has 300 amino acids, and the molecular weight of the protein is 33.8kDa. When the KNAT7 is specifically expressed in cotton fibres, a great deal of transcription products of the KNAT7 can be accumulated in fibres at the later stage of cell elongation and the early stage of secondary wall thickening. The expression of the KNAT7 gene in yeast can promote the longitudinal elongation of host cells, and a transgenic arabidopsis analysis shows that the synthesis of secondary wall components is affected by the KNAT7 to a certain degree. The experimental result proves that the KNAT7 plays an important role in the processes of cotton fibre elongation and secondary wall thickening, and the gene has important application value in improving the quality of cotton fibres and increasing the yield of cotton.

Description

Technical field: [0001] The invention relates to cotton genes, specifically the cotton fiber cell-specific expression and functional identification of a new member gene KNAT7 in the cotton homeobox (HB) family, and is further applied to the improvement of cotton fiber quality. Background technique: [0002] Cotton is one of the most important economic crops in my country, and cotton fiber is widely used in the textile industry. With the rapid development of the textile industry and the continuous improvement of people's living standards, the textile industry and its related industries have higher and higher requirements for the quality of cotton fibers. my country's cotton output ranks first in the world, but the quality of cotton fiber is poor and cannot be used as raw materials for the production of some high-value-added high-quality textile products. Therefore, my country needs to import more than 1 million tons of high-quality cotton every year, resulting in a backlog o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/68C12Q1/02A01H5/00
CPCY02A40/10
Inventor 李学宝龚思颖孙翔赵黎莉
Owner HUAZHONG NORMAL UNIV
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