Protein related to flowering time of plants and applications of protein

A technology of flowering time and plant, applied in protein and its application field

Active Publication Date: 2020-09-04
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, expansins related to plant flowering time have not been reported yet.

Method used

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  • Protein related to flowering time of plants and applications of protein
  • Protein related to flowering time of plants and applications of protein
  • Protein related to flowering time of plants and applications of protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Cloning of embodiment 1 early flowering gene PtoEXPB3

[0070] The inventors of the present invention isolated and cloned an early flower-related gene PtoEXPB3 from Populus tomentosa TC1521, as shown in sequence 1 of the sequence listing, and named its encoded protein as PtoEXPB3 protein, as shown in sequence 2 of the sequence listing.

[0071] The specific cloning method is as follows:

[0072] Utilize the Trizol method (Invitrogen, USA) to extract the total RNA of Populus tomentosa TC1521 leaves, reverse transcribe into cDNA, use cDNA as template, and use primer pair (PtoEXPB3F:5'-CCAGAAAACTGTCGTCAAAGG-3';

[0073] PtoEXPB3R: 5'-GCTTGTTTCTCTCACGTGGA-3') was used as a primer, and KODplus DNA polymerase was used to amplify the gene PtoEXPB3. The amplification program was: 94°C for 5min; 94°C for 30s, 55°C for 40s, 72°C for 1min, a total of 35 cycles. The amplified products were separated by 1.5% agarose gel electrophoresis, and the electrophoresis results were as foll...

Embodiment 2

[0075] Example 2 Obtaining and identification of Nicotiana benthamiana transformed with PtoEXPB3 gene

[0076] 1. Construction of recombinant expression vector

[0077] With the recombinant vector P-Blunt-PtoEXPB3 in embodiment 1 as template, with PtoEXPB3F (5'-CCC AAGCTT ATGCAGCTCTTGGGGTTAC-3', the underline is the restriction site of HindIII) and PtoEXPB3R (5'-TGC TCTAGA TTAATGGAAGAAATTGAGCCTAGAGG-3', the underline is the restriction site of XbaI) as a primer pair, and the high-fidelity enzyme KOD is used to amplify and recover the PCR product. The PCR product and the PEZR(K)-LC expression vector plasmid were respectively digested with Hind III and Xba I restriction endonucleases, and the digested PEZR(K)-LC vector and target gene fragment were recovered to obtain the recombinant vector PEZR( K)-LC-PtoEXPB3 (structural schematic diagram as image 3 shown), the recombinant vector PEZR (K)-LC-PtoEXPB3 is that the sequence between the Hind III and Xba I recognition sites o...

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Abstract

The invention relates to protein related to the flowering time of plants and applications of the protein. The protein is shown as follows: A1) the amino acid sequence of the protein is shown as a sequence 2 in a sequence table; A2) the protein obtained by substituting and/or deleting and/or adding one or more amino acid residues to the amino acid sequence shown as the sequence 2 in the sequence table has more than 90% identity with the protein shown in the A1) and is related to the flowering time of plants; and A3) the fusion protein is obtained by linking the N-terminal or/and C-terminal of the A1) or the A2) and protein tags. An extended protein gene PtoEXPB3 is obtained from populus tomentosa. The gene is introduced in tobacco; and compared with wild type tobacco, the flowering time oftransgenic tobacco flowers can be advanced by 21 days, so that the PtoEXPB3 can have the function of early flowering and can be taken as an important gene resource of the molecular breeding of ornamental plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein related to plant flowering time and its application. Background technique [0002] The early flowering phenomenon of plants (that is, early budding and flowering) is related to genes, and some early flowering genes have been identified in different plants, and have had a positive impact in production practice. For example, Smykal et al. (2007) cloned the genes NtSOC1 and NtFUL related to flowering time from tobacco, which are related to photoperiod characteristics, and overexpression can cause early flowering of tobacco. Li Shuang et al. (2014) introduced the PsnAP1 gene of Populus nigra in tobacco, and the transgenic plants flowered earlier. Wang (2012) found that overexpression of PtGT1 gene in tobacco plants showed early flowering. An Xinmin et al. (An et al., 2011) showed that the PtAP3 gene of Populus tomentosa plays an important role in inducing early flowering of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/827
Inventor 徐吉臣张皓尹鹏刘晓徐筱
Owner BEIJING FORESTRY UNIVERSITY
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