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Chinese cabbage BcpFLC3 protein encoding array

A coding sequence, Chinese cabbage technology, applied in the field of genetic engineering, can solve problems that have not yet been discovered

Inactive Publication Date: 2004-02-04
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the analysis of prior art literature, although "The Plant Journal, 2001, 28: 545-553" has fully affirmed the function of the delayed flowering of the FLC gene, especially BnFLC 1-5 And the determination of AtFLC gene function has made great progress in the application of people in delaying flowering, but has not found the report that has close connection literature with subject of the present invention so far

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Cloning of Bolting Resistance Gene in Chinese Cabbage

[0057] 1. Organization separation (isolation)

[0058] Chinese cabbage (variety "Chun Dawang") is purchased from the market. The Chinese cabbage is placed at 28°C for 24 hours to germinate, and then sown in a greenhouse. When the number of Chinese cabbage leaves is 4-5, DNA or RNA is prepared.

[0059] 2. RNA isolation (RNA isolation)

[0060] Take part of the tissue, grind it with a mortar, add a 1.5mL EP tube containing the lysis solution, shake it enough, and then move it into a glass homogenizer. After homogenization, transfer to a 1.5mL EP tube to extract total RNA (TRIzol Reagents, GIBCO BRL, USA). Use formaldehyde denaturing gel electrophoresis to identify the total RNA quality, and then measure the RNA content on a spectrophotometer.

[0061] 3. Cloning of Full-length cDNA

[0062] According to the amino acid conservative sequence of the Arabidopsis thaliana suppressing flowering gene, using the principle of ho...

Embodiment 2

[0074] Sequence Information and Homology Analysis of Chinese Cabbage Bolting Resistance Related Genes

[0075]The length of the new Chinese cabbage bolting-resistant full-length cDNA of the present invention is 851 bp, and the detailed sequence is shown in SEQ ID NO. 3, in which the open reading frame is located at nucleotides 55-645 (591 nucleotides). According to the full-length cDNA, the bolting-resistant amino acid sequence of Chinese cabbage was deduced, with a total of 197 amino acid residues, a molecular weight of 21644.98 Daltons, and an isoelectric point (pI) of 9.25. See SEQ ID NO.3. for detailed sequence.

[0076] The full-length cDNA sequence related to bolting resistance of Chinese cabbage and its encoded protein were used BLAST program to carry out nucleotide and protein in Non-redundant GenBank +EMBL+DDBJ+PDB and Non-redundant GenBank CDStranslations +PDB+ SwissProt+Superdate+PIR database Homology search revealed that it is 98% identical to the flowering-suppressing...

Embodiment 3

[0078] Chinese cabbage bolting-resistance-related protein or polypeptide in Arabidopsis thaliana for eukaryotic expression and bolting identification of transgenic plants

[0079] Construction of an expression vector containing the target gene (Chinese cabbage BcpFLC3 gene)

[0080] According to the full-length sequence related to bolting resistance of Chinese cabbage (SEQ ID NO.3), primers were designed to amplify the complete coding reading frame, and restriction endonuclease sites were introduced on the upstream and downstream primers (this can be selected Depending on the vector) in order to construct an expression vector. Using the amplified product obtained in Example 1 as a template, after PCR amplification, the Chinese cabbage bolting resistance gene cDNA was cloned into an intermediate vector (such as pBluescript), and then cloned into a binary expression vector (such as pBI121 and improved pCAMBIA2300) , Under the premise of ensuring the correct reading frame, the identi...

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Abstract

A coding sequence of Chinese cabbage BcpFLC3 protein and the nucleotide sequence for coding the polypeptide with the activity of said BcpFLC3 protein are disclosed, which can suppress the bolting or delay blossom, so having great application value in Chinese cabbage, flower, etc.

Description

Technical field [0001] The present invention relates to a protein coding sequence, especially a Chinese cabbage BcpFLC3 protein coding sequence, which belongs to the field of genetic engineering. Background technique [0002] In recent years, there have been more and more researches on genes and their proteins that control flowering time in Cruciferae. The MADS box gene belongs to an ancient gene family, exists in the entire eukaryotic world, and its biological function is to regulate cell differentiation and development. The protein products of the MADS gene family are transcription factors, which have the ability to regulate the transcription of other genes. The first four member genes identified are MCM1, AGAMOU, DEFICIENS, and SRF. The first letters of these genes constitute MADS. In terms of gene structure, the members of the MADS gene family all contain a highly conserved sequence called MADS box (MADSbox), which encodes about 50 amino acids, and is related to the recogniti...

Claims

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Application Information

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IPC IPC(8): C07H21/00C07K14/415C12N15/29C12N15/62C12N15/82
Inventor 唐克轩李柱刚赵凌侠孙小芬开国银
Owner SHANGHAI JIAO TONG UNIV
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