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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 