Catalpa bungei CabuAP3 protein as well as encoding gene and application thereof

A technology that encodes genes and proteins, applied in the field of molecular biology, can solve problems such as poor sequence homology, highly unconserved sequences, and difficulty in obtaining AP3 homologous genes, and achieve good application prospects

Inactive Publication Date: 2015-12-02
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows us to study how certain proteins called Cabou AP 3 (CAB) affect plants' growth or behavior during different stages such as flowering, seed germination, nutrients absorption, photosynthesis, etc., which could lead to improved agricultural production efficiency through increased yielding from crops grown under specific conditions.

Problems solved by technology

This patented technical problem addressed in this patents relates to studying how certain types of DNA have various functions that control their own biological processes like cell division and embryogenesis during seedling stages.

Method used

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  • Catalpa bungei CabuAP3 protein as well as encoding gene and application thereof
  • Catalpa bungei CabuAP3 protein as well as encoding gene and application thereof
  • Catalpa bungei CabuAP3 protein as well as encoding gene and application thereof

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Embodiment 1

[0030] The cloning of embodiment 1 a Chinese catalpa CabuAP3 gene cDNA sequence

[0031] 1. Extraction of totalRNA from flower buds of Chinese catalpa

[0032] Fresh flower buds with a length of about 5 mm were collected from Catalpa, put into cryopreservation tubes, immediately put into liquid nitrogen for quick freezing for 3 hours, and then put into -80°C ultra-low temperature freezer for later use. Use the RNA extraction kit to extract the total RNA of Chinese catalpa flower buds: take out the material of Chinese catalpa flower buds from the -80°C ultra-low temperature freezer, put them into a mortar with 2mL RLT lysate and 100μL PLANTaid, and grind them fully at room temperature; Transfer to a 2mL eppendorf tube, centrifuge at 13000rpm for 10min, absorb 500μL supernatant, and transfer to a new 2mL centrifuge tube; add 250μL absolute alcohol to the supernatant, blow and mix well; transfer the above liquid to an adsorption column, and Put the adsorption column into the col...

Embodiment 2

[0041] Example 2 The protein sequence encoded by a Chinese catalpa CabuAP3 gene

[0042] Using the primer5 software, the full-length cDNA sequence of the Catalpa CabuAP3 gene was translated into a protein sequence (SEQ ID No.1), and according to the characteristics of the MADS-box gene, the CabuAP3 protein sequence of the Catalpa tree was translated into the M region, I region, K region, and C region And the division of the 2 motifs PI-derived motif and euAP3 motif contained in the C region ( figure 2 ).

Embodiment 3

[0043] The expression pattern analysis of embodiment 3 Chinese catalpa CabuAP3 gene

[0044] According to the full-length cDNA sequence of Catalpa cabuAP3 gene, oligo6.0 software was used to design primers RTAP3F:5'-TACATCAGCCCCACTATAACGA-3' and RTAP3F:5'-TTATTCAAGCAAAGCAAACGTG-3' for semi-quantitative experiments. Use PCR to detect its specificity, and it can only be used under the premise of ensuring PCR specific amplification. The catalpa actin gene was used as the positive control of the semi-quantitative test, and the primers of the actin gene were CabuactinF: 5'-TCACACTGGTGTGATGGTTG-3' and CabuactinR: 5'-AGTTCTTCTCCACAGCAGAG-3'. The detection band brightness was adjusted by the proportion of the first-strand cDNA template amount in young leaves, sepals, petals, stamens and pistils. Semi-quantitative PCR reaction program: 94°C for 4min; 94°C for 30s, 57°C for 30s, 72°C for 30s, 25 cycles; 72°C for 10min. After the reaction, electrophoresis detection was carried out with...

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Abstract

The invention relates to the field of molecular biology, and in particular to catalpa bungei CabuAP3 protein as well as an encoding gene and application thereof. The amino acid sequence of the catalpa bungei CabuAP3 protein is as shown in SEQ ID No.1; the sequence (CDS) of the encoding gene is as shown in SEQ ID No.2; the whole length of the cDNA sequence of the catalpa bungei CabuAP3 gene is as shown in SEQ ID No.3. The catalpa bungei CabuAP3 gene provided by the invention is only expressed in petals and stamens of catalpa bungei, and is not expressed in leaves, sepal or pistil. A CabuAP3 gene expression vector is transferred into mutants of wild arabidopsis and arabidopsis ap3-3, the result shows that the transgenic wild arabidopsis has one extra petal, which indicates that the CabuAP3 gene has the function of regulating and controlling increase of the number of petals of the transgenic wild arabidopsis, and the transgenic wild arabidopsis ap3-3 mutant has a filament structure. The plant variation material obtained from the catalpa bungei CabuAP3 gene expression vector can be applied to plant breeding and view admire, and has very good application prospect.

Description

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Claims

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Application Information

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Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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