Rapid cloning method and special primers for lfy/flo homologous fragments of plant flowering-related genes

A cloning method and plant technology, applied in the direction of plant genetic improvement, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of high cost and long time consumption, and achieve the effect of fast method and wide variety of plants

Inactive Publication Date: 2011-12-07
SOUTHWEST FORESTRY UNIVERSITY
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Problems solved by technology

Although the above methods have been applied to a certain extent, th

Method used

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  • Rapid cloning method and special primers for lfy/flo homologous fragments of plant flowering-related genes
  • Rapid cloning method and special primers for lfy/flo homologous fragments of plant flowering-related genes
  • Rapid cloning method and special primers for lfy/flo homologous fragments of plant flowering-related genes

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Embodiment 1

[0033] (1) Design and synthesis of special primers, namely degenerate primers LYF and LYR

[0034] According to the reported LFY / FLO Degenerate primers LYF and LYR designed for homologous gene sequences, the nucleotide sequence of primer LYF is 5'- TAYATIAAYAARCCIAARATG -3' (as shown in SEQ ID NO:1), the nucleotide sequence of primer LYR is 5' -ARIYKIGTIGGIACRTACCA-3' (ie, as shown in SEQ ID NO: 2), the expected amplified target product is 236bp.

[0035] The above special primers were synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.

[0036] (2) Use the above-mentioned special primers for the above 14 kinds of plants LFY / FLO The homologous fragments were amplified by PCR, and the steps were as follows:

[0037] ① Collection of samples

[0038] The 14 kinds of plant samples were taken from various parks in Kunming City, Yunnan Province.

[0039] ② DNA extraction

[0040] DNA was extracted from plant tissues using the Quick Plant Genomic DNA K...

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Abstract

The invention relates to a rapid cloning method of plant flowering related gene LFY/FLO homologous fragments and special primers thereof, and relates to the technical field of rapid isolation and cloning of plant homologous genes. The method is characterized in that the used special primer LYF has a nucleotide sequence of 5'-TAYATIAAYAARCCIAARATG-3', and LYR has a nucleotide sequence of 5'-ARIYKIGTIGGIACRTACCA-3'; the primer concentration is 25 muM, and the annealing temperature is 44 DEG C. The method of the invention can clone to obtain homologous fragments of LFY/FLO genes of plants from bryophytes to spermatophytes, and has a wide range of the kinds of applicable plants; the potential problem that a gene fragment is difficult to be amplified due to no expression or low expression of homologous fragments of the LFY/FLO gene when cDNA is used as a template is well avoided; and the method is rapid, simple, economical, and effective.

Description

technical field [0001] The present invention relates to the technical field of rapid separation and cloning of plant homologous genes, in particular to the process from bryophytes to seed plants LFY / FLO A rapid separation method for gene homologous fragments and its special primers. Background technique [0002] The flowering process of plants is the result of the coordination of the external environment and internal genetic factors. In the flowering regulatory network, LFY The gene is the hub of GA signal and photoperiod-induced signal transduction. It is an essential gene that determines the formation of floral meristem and plays an important role in activating the activity of floral organ genes. LFY / FLO Homologous genes are present in both flowering and non-flowering higher plants. LFY / FLO Homologous genes, as characteristic genes of floral meristems, are important elements that determine the transition from vegetative growth to reproductive growth, and contro...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/29C12N15/10
Inventor 鄢波于丽霞汤晓倩姚有林樊国盛
Owner SOUTHWEST FORESTRY UNIVERSITY
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