Chinese medicine composition anti-wrinkle multi-function liquid, its preparation and application

A gene and protein technology, which is applied to plant resistance-related protein ATSAR42 and its encoding gene and application fields, can solve the problems of increasing salicylic acid, increasing the synthesis level of salicylic acid, which have not yet been found, and achieves the effect of enhancing disease resistance.

Active Publication Date: 2013-06-12
FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD
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AI Technical Summary

Problems solved by technology

So far, it has only been found that ETHYLENEINSENSITIVE3 (EIN3) and EIN3-LIKE1 can directly bind to the ICS1 promoter of the key SA biosynthesis gene, and negatively regulate the synthesis level of SA. It has not been found that ICS1 gene expression can be promoted by positively inducing the ICS1 promoter. proteins that increase the level of salicylic acid synthesis
In addition, studies have found that SAR Deficient 1 (SARD1) and CBP60g proteins are important factors in inducing ICS1 gene expression and SA synthesis, but failed to find more positive induction of ICS1 promoter to promote ICS1 gene expression, thereby increasing the level of salicylic acid synthesis protein

Method used

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  • Chinese medicine composition anti-wrinkle multi-function liquid, its preparation and application
  • Chinese medicine composition anti-wrinkle multi-function liquid, its preparation and application
  • Chinese medicine composition anti-wrinkle multi-function liquid, its preparation and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, the acquisition of ATSAR42 protein and its coding gene

[0062] 1. According to the existing NCBI database and literature, set a pair of primers as follows:

[0063] F1 (forward primer): 5'-ccg ctcgag tttgctttgtatgctcaacaatt-3' (XhoI restriction recognition sequence is underlined);

[0064] R1 (reverse primer): 5'-cg ggatcc atctaggagccaactctgtg-3' (the BamHI restriction recognition sequence is underlined).

[0065] 2. Genomic DNA was extracted from leaves of Arabidopsis thaliana ecotype Columbia.

[0066] 3. Using genomic DNA as a template, use the primer pair designed in step 1, and perform PCR amplification with PrimeSTAR HS high-fidelity enzyme from TaKaRa Company.

[0067] 4. The PCR amplification product is sequenced, as shown in sequence 2 of the sequence listing.

[0068] The protein shown in Sequence 1 of the sequence listing is named ATSAR42 protein. The gene encoding the ATSAR42 protein is named as the ATSAR42 gene, and its genomic DNA is...

Embodiment 2

[0069] Embodiment 2, the cloning of each gene and the construction of its recombinant expression vector

[0070] 1. Acquisition of ATSAR42 gene and construction of recombinant plasmid 326-ATSAR42-FLAG

[0071] 1. Genomic DNA was extracted from leaves of Arabidopsis thaliana ecotype Columbia.

[0072] 2. Using the genomic DNA in step 1 as a template, use the primer pair composed of F1 and R1 to carry out PCR amplification under the action of TaKaRa's PrimeSTAR HS high-fidelity enzyme to obtain PCR amplification products. The agarose gel electrophoresis of the PCR amplification product is shown in figure 1 (M represents a nucleotide marker, which is DL2000 from TaKaRa Company).

[0073] 3. The PCR amplified product of step 2 was double-digested with restriction endonucleases XhoI and BamH I, and the digested product was recovered.

[0074] 4. Digest the 326-FLAG expression vector with restriction endonucleases XhoI and BamH I, and recover the vector backbone of about 3827bp.

...

Embodiment 3

[0096] Embodiment 3, the application of ATSAR42 gene in inducing ICS1 gene promoter (ProAtICS1) to start gene expression

[0097] The recombinant plasmid 326-ATSAR42-FLAG and recombinant plasmid 326-Pro constructed in Example 2 AtICS1 ::GFP co-transformed Arabidopsis protoplasts (recombinant plasmid 326-T 7 -FLC is used as a negative control of the recombinant plasmid 326-ATSAR42-FLAG; the 326-FLAG expression vector is used as another negative control of the recombinant plasmid 326-ATSAR42-FLAG), the specific steps are as follows:

[0098] 1. Germinate Colombian ecotype Arabidopsis seeds on MS medium, transplant them into soil when the roots grow to 1-3 cm, and cultivate them in a greenhouse at 23° C. (12 hours of light per day, light intensity of 150 μE).

[0099] 2. Add 20ml of double distilled water to a 90mm petri dish, then add 1.82g of D-mannitol and dissolve it.

[0100] 3. Take the unbolted leaves (about 90 pieces) that have been cultured for 4 weeks in step 1, cut t...

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Abstract

The invention discloses a Chinese medicine composition anti-wrinkle multi-function liquid, its preparation and application. The anti-wrinkle multi-function liquid comprises the following ingredients: 11-13 weight portions of the root of kudzu vine, 12-14 weight portions of rehmannia glutinosa, 7-9 weight portions of gingko, 11-13 weight portions of dark plum, 12-14 weight portions of leonurus, 4-6 weight portions of radix salvia miltiorrhizae, 6-8 weight portions of liquorice, 7-9 weight portions of Ligusticum wallichii, 6-8 weight portions of Angelica sinensis, 5-7 weight portions of Semen Cuscutae, 3-5 weight portions of glossy ganoderma, 4-6 weight portions of rhizoma cyperi, and 8-10 weight portions of ginseng. The multi-function liquid is light yellow and tasteless. The trade name ofthe multi-function liquid is aromlotus Chinese medicine composition anti-wrinkle multi-function liquid. The direct use of the multi-function liquid on the skin can effectively remove wrinkles, and can be absorbed by the skin effectively, so that the effects of skin care and skin beauty are achieved. According to the invention, no chemical additives are added in the multi-function liquid, and the multi-function liquid uses all natural extracts, so that the effects are realized depending on the Chinese medicine extracts completely; and the natural extracts are obtained by mixed extraction according to Chinese medicine formula, so that the characteristics of the compatibility of Chinese medicines are really shown.

Description

technical field [0001] The invention relates to a plant resistance related protein ATSAR42 and its coding gene and application. Background technique [0002] Plants often suffer from various diseases in the process of cultivation and production. Some diseases pose a great threat to plant growth and seriously damage the yield and quality of crops. For plant diseases, at present, the main measures are to control them with drugs. Recently, people have begun to use abiotic inducers to induce plant disease resistance, and have been widely used in many important crops such as tobacco, potato, tomato, cucumber, bean, and rice. Although the effect of this measure is ideal, it is costly and pollutes the environment. Therefore, there is an urgent need to develop new biological means to improve the plant's own disease resistance. [0003] Systemic acquired resistance is a plant defense response that is induced when plants are attacked by pathogens and pests and can quickly spread to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415
Inventor 金京波蔡斌林晓莉
Owner FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD
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