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Cotton ghuxs3 gene, its encoded protein and its application

A technology of gene coding and protein, applied in the field of genetic engineering, to achieve the effect of improving the quality and properties of cotton fiber

Inactive Publication Date: 2011-11-30
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no related report on this gene in cotton

Method used

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  • Cotton ghuxs3 gene, its encoded protein and its application
  • Cotton ghuxs3 gene, its encoded protein and its application
  • Cotton ghuxs3 gene, its encoded protein and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Cloning of embodiment 1 cotton GhUXS3 gene

[0030] (1) Firstly, using the coding sequence of Arabidopsis four uridine diphosphate glucuronate decarboxylase (AT3g62830, AT5g59290, AT2g47650 and AT3g46440), search for EST sequences with high similarity in the NCBI cotton expressed sequence tag database by using tBLASTN, The retrieved EST sequences (CO494958, DW504337 and DW515118) were electronically assembled using DNASTAR software to obtain the spliced ​​sequence GhUXS3 with a complete open reading frame.

[0031] (2) Design specific primers according to the spliced ​​sequence, and simultaneously introduce SacI and SalI restriction sites (underlined parts) into the upstream and downstream primers, respectively. The upstream and downstream primers of the GhUXS3 gene are:

[0032] Upstream primer: 5'-CTA GAGCTC ATGGCGACAGATTCATC-3',

[0033] Downstream primer: 5'-GCG GTC GAC TCACTCTTTTAGAGATTCC-3';

[0034] (3) Using the designed specific primers to amplify the cD...

Embodiment 2

[0036] Example 2 Similarity Analysis of the Amino Acid Sequences of the Cotton GhUXS3 Gene and 5 Arabidopsis UDP-Glucuronate Decarboxylase Gene-Encoded Proteins

[0037] Using BLAST (bl2seq) to analyze the similarity of the amino acid sequences of the proteins encoded by the cotton GhUXS3 gene and five Arabidopsis UDP-glucuronate decarboxylase genes, the results showed that the cotton GhUXS3 gene and the Arabidopsis UDP-glucuronate decarboxylase gene The similarity between the amino acid sequences of the encoded proteins was distributed between 66% and 91%. It shows that the cotton GhUXS3 gene belongs to the UDP-glucuronate decarboxylase family.

[0038] Using DNAMAN software to perform multiple sequence comparison analysis on the above protein sequences, the results are as follows figure 2 shown. The protein encoded by cotton GhUXS3 gene has the conserved domains GxxGxxG, YxxxK and Ser of UDP-glucuronate decarboxylase protein. It also shows that the cotton GhUXS3 gene bel...

Embodiment 3

[0039] Cloning of embodiment 3 cotton GhUXS3 gene 5' and 3' untranslated region (UTR) sequence

[0040] Based on the open reading frame sequence of the cotton GhUXS3 gene, 5'RACE and 3'RACE primers were designed according to the requirements of the Clontech RACE kit. The specific primer sequences are as follows:

[0041] 5' RACE primers:

[0042] GSP: 5′-TCGCTTGGCAAGTCCCCAGCATGT-3′

[0043] NGSP: 5′-ATTGGAGAAGCTGGACACGCAAGGT-3′

[0044] 3' RACE primers:

[0045] GSP: 5′-CACTTCGCGGTGAACCACTAACGGT-3′

[0046] NGSP: 5′-GCTAGGTTGGGAGCCGAAAGTCAAGT-3′

[0047] Using cotton fiber cDNA 21 days after flowering of upland cotton "Zhongmiansuo 8" and sea island cotton "Pima90-53" as materials, the 5' and 3' untranslated regions were amplified according to the operation steps of the Clontech RACE kit , to obtain the target band, GhUXS35'RACE length 452bp, 3'RACE length 488bp ( image 3 ). The PCR product was sequenced, and the final analysis showed that the length of GhUXS35'UTR was...

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Abstract

The invention provides a cotton uridine diphosphate glucuronate decarboxylase (GhUXS3) gene and a protein encoded by the gene. A nucleotide sequence and an amino acid sequence of the gene are shown as SEQID No.1 and SEQ ID No.2 respectively. GhUXS3 is a key enzyme for the formation of non-cellulose substances in the developmental process of cotton fibers, and affects the quality of the cotton fibers, so the novel candidate gene is provided for genetic engineering of quality improvement of the cotton fibers; and the GhUXS3 gene can lay a foundation for further transforming cotton so as to improve the quality of the cotton fibers, and has greater economic benefit and application value.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the cotton GhUXS3 gene, its encoded protein and its application. Background technique [0002] The quality of cotton fiber is determined by the development of cotton fiber. Cotton fiber development goes through four stages: fibroblast differentiation and protrusion, fiber cell elongation (primary wall formation stage), secondary wall thickening, and dehydration maturity. The primary wall formation period and the secondary wall thickening period are the key periods for the formation of cotton fiber quality. Among them, the primary wall formation period is mainly the synthesis of non-cellulose substances such as xyloglucan, xylan and pectin, which determine the length of cotton fibers. The secondary wall thickening stage is a process of mass deposition of cellulose, which is directly related to fiber strength. Therefore, to clarify the genes that affect the formation of cotton...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12N15/60C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
Inventor 马峙英潘玉欣张冬梅王省芬张桂寅
Owner HEBEI AGRICULTURAL UNIV.
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