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Promoter and application thereof

A technology of promoters and DNA molecules, applied in the field of promoters, can solve problems such as differences and achieve the effect of cultivation and promotion

Active Publication Date: 2014-12-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression of HGTP in hair follicles has strict spatiotemporal expression specificity, but there are differences among different species

Method used

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  • Promoter and application thereof
  • Promoter and application thereof
  • Promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, discovery of promoter

[0022] On the basis of the KAP6.1 gene and its known promoter sequence, it was found that the new promoter obtained by extending the known promoter sequence to the 5' flanking region by 481bp has a higher activity of promoting the expression of the target gene.

[0023] The promoter shown in the 488th to 1529th nucleotides from the 5' end of Sequence 1 in the sequence listing is named as promoter A (known promoter).

[0024] The promoter shown in the 7th to 1529th nucleotides from the 5' end of Sequence 1 in the sequence listing is named as promoter B (promoter provided by the present invention).

Embodiment 2

[0025] Example 2, functional verification of the promoter

[0026] F1: 5'-cta gctagc CTGCAGTTCATGGGGTCAC-3';

[0027] F2: 5'-cta gctagc TCCGTTTTACTAAGAAGCATTTT-3';

[0028] R: 5'-ccg ctcgag GGTGTTGCTTGTTGAGGTTG-3'.

[0029] 1. Construction of recombinant plasmid A

[0030] 1. Extract the genome DNA of Aohan fine-wool sheep.

[0031] 2. Using the genomic DNA extracted in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R to obtain a PCR amplification product. The agarose gel electrophoresis of the PCR amplification product is shown in figure 1 (M is a 2kb marker, 1, 2, and 3 are PCR amplification products).

[0032] PCR reaction conditions: 95°C, 5min; 35 cycles of 95°C for 30s, 60°C for 30s, 72°C for 1.5min; 72°C for 7min.

[0033] 3. Digest the PCR amplified product in step 2 with restriction endonucleases NheI and XhoI, and recover the digested product.

[0034] 4. The pGL3-Basic vector was double-digested with restriction en...

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Abstract

The invention discloses a promoter and application thereof. The promoter is acquired from Aohan merino sheep and is a deoxyribonucleic acid (DNA) molecule from (1) a DNA molecule shown by a sequence 1 of a sequence table, (2) a DNA molecule which is hybridized with the DNA sequence limited in (1) under strict conditions and has a promoter function, and (3) a DNA molecule which has more than 90 percent of homology with the DNA sequence limited in (1) and has a promoter function. Compared with an existing promoter, the promoter disclosed by the invention has higher activity for promoting the expression of a target gene, and can meet the expression requirement of the target gene in a sheep skin tissue as well as the requirement on the high-efficiency specific expression of the target gene in a hair follicle tissue. The promoter not only can be used for researching a control mechanism for the gene expression in hair fineness and curvedness and hair follicle development, but also can promote the breeding of the transgenic ultrafine-hair merino sheep.

Description

technical field [0001] The present invention relates to a promoter and its application. Background technique [0002] Today's wool textiles are developing in the direction of light, soft and high-grade, which makes the wool textile industry have higher and higher requirements for the comprehensive quality of fine wool sheep. Over the past 20 years, the fineness of Australian wool has undergone great changes. At present, more than 1 / 3 of wool belongs to superfine wool, and this trend is still developing. In order to meet the market demand, my country began to select and breed Chinese Merino superfine wool sheep in the 1980s. But so far the fineness of ultra-fine wool cultivated in my country has not yet reached the international standard, and the output cannot be produced in batches, so there is a large gap between supply and demand. [0003] In the study of breeding transgenic superfine wool Merino sheep, it is urgent to obtain skin hair follicle tissue-specific expression...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/21C12N1/19C12N1/15C12N5/10
Inventor 邓学梅杨祖崔凯
Owner CHINA AGRI UNIV