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Purple sweet potato ultraviolet receptor UVR8 gene and cloning method thereof

A cloning method and ultraviolet light technology, applied in genetic engineering, plant genetic improvement, chemical instruments and methods, etc., can solve the problem of lack of sweet potato UVR8 gene, etc., and achieve the effect of simple method, saving time and cost, and high success rate

Inactive Publication Date: 2018-06-01
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

[0004] At present, there is no report on the sweet potato UVR8 gene, and it is quite necessary to clone the full sequence of the sweet potato UVR8 gene

Method used

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  • Purple sweet potato ultraviolet receptor UVR8 gene and cloning method thereof
  • Purple sweet potato ultraviolet receptor UVR8 gene and cloning method thereof
  • Purple sweet potato ultraviolet receptor UVR8 gene and cloning method thereof

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Embodiment Construction

[0032] 1 Materials and methods

[0033] 1.1 Test material

[0034] Fu Caishu No. 23, (from the sweet potato germplasm resource nursery of the Crop Research Institute of Fujian Academy of Agricultural Sciences, literature source: Li Guoliang et al., Effects of light and temperature on anthocyanin synthesis and gene expression of related enzymes in leaf vegetable sweet potato leaves. 2017 ) is a new variety of sweet potato for leaf vegetables bred by the Crop Research Institute of Fujian Academy of Agricultural Sciences, and it is planted in the scientific research and breeding base. Sweet potato leaves grown normally in the field were taken, quick-frozen in liquid nitrogen, and stored in a -80°C refrigerator.

[0035] TransStart Top Green qPCR SuperMix was purchased from Beijing Quanshijin Biotechnology Co., Ltd. RNA extraction reagents, D2000marker, pMD18-T vector and RACE kit were purchased from Takara Company. OligodT18 primers were purchased from Treasure Bioengineering ...

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Abstract

The invention provides a purple sweet potato ultraviolet receptor UVR8 gene and a cloning method thereof. A nucleotide sequence of the purple sweet potato ultraviolet receptor UVR8 gene is shown as SEQ NO. 4; the cloning method of the purple sweet potato ultraviolet receptor UVR8 gene comprises the following steps: (1) synthesizing a cDNA (complementary Deoxyribonucleic Acid) first chain; (2) carrying out homologous cloning on a UVR8 conserved sequence; (3) amplifying a 3' end of UVR8; (4) amplifying a 5' end of the UVR8; (5) splicing the sequence. By adopting the method provided by the invention, species with a relatively close genetic relationship are not needed and the cloning of a core fragment can be finished through designing various merged basic groups only if a homologous gene exists; the method is also applicable to gene cloning of other plants. In 5' end cloning, a process that a phosphoric acid group of mRNA (messenger Ribonucleic Acid) is removed through phosphorylase and ligase is connected with a connector linker primer is not needed; howeverbut, 3 to 5 C are added at the 3' end in an RNA reverse transcription process according to properties of M-MuLV reverse transcriptase; then 3 to 5 G are added at the tail end of a designed primer and are matchedfoe matching with the designed primerC and the reverse transcription process is finished. The method is simple and convenient, the time and cost are saved and the success rate is relatively high.

Description

technical field [0001] The invention relates to a photoreceptor UVR8 gene and a cloning method thereof, in particular to a sweet potato ultraviolet light receptor UVR8 gene and a cloning method thereof. Background technique [0002] Light is one of the most important environmental factors affecting plant growth and development in nature. It can not only provide radiation energy for photosynthesis of plants, but also act as a signal molecule to regulate its growth and development process (Gao Rongfu et al., 2002). The photoreceptors in plants can sense light and generate signal transmission. The plant photoreceptors that have been discovered so far include phytochromes (phyA-E) that feel red light and far-red light, and cryptogams that feel blue light / UV-A. Pigments (cryptochromes, cry1 and cry2) and phototropins (phototropins, phot1 and phot2) and the receptor UVR8 (UV RESISTANCE LOCUS 8) for ultraviolet light (UV-B) (Quail, 2002; Banerjee and Batschauer, 2005; Jenkins , 20...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12Q1/6806
CPCC07K14/415C12Q1/6806
Inventor 李国良邱永祥邱思鑫林赵淼许泳清刘中华张鸿李华伟纪荣昌罗文彬汤浩
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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