Human KLF7 gene promoter as well as construction method and application thereof

A promoter, gene technology, applied in the field of genetic engineering

Active Publication Date: 2021-04-16
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there have been no reports on the promoters for the expression of various human KLF7 transcripts. Confirming the promoter of the human KLF7 gene has appli

Method used

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  • Human KLF7 gene promoter as well as construction method and application thereof
  • Human KLF7 gene promoter as well as construction method and application thereof
  • Human KLF7 gene promoter as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of embodiment 1 human KLF7 gene promoter

[0034] 1. Human sperm genome DNA extraction

[0035] 1.1 Reagent configuration

[0036] 1) Phenol: chloroform: isoamyl alcohol = 25:24:1 (prepared according to volume ratio)

[0037] 2) Chloroform: isoamyl alcohol = 24:1 (prepared according to volume ratio)

[0038] 3) 20mg / mL proteinase K (proteinase K): Add 200mg of proteinase K to 9.5mL of double-distilled water, shake gently until the proteinase K is completely dissolved, dilute to 10ml, then aliquot and store at -20°C.

[0039] 4) 10% SDS (sodium dodecyl sulfate): Weigh 100 g of SDS and slowly transfer it to a beaker of 0.9 L of double distilled water, heat to 68° C. and stir with a magnetic stirrer until completely dissolved. If necessary, adjust the pH value to 7.2 with 1mol / L NaOH, dilute to 1L with double-distilled water, and store at room temperature without sterilization.

[0040] 5) STE preparation

[0041] 10mmol / L Tris-HCl (PH8.0)

[0042] 0.1mol...

Embodiment 2

[0085] Analysis of Human KLF7 Gene Promoter Activity

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PUM

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Abstract

The invention discloses a human KLF7 gene promoter as well as a construction method and application thereof, and belongs to the technical field of gene engineering. The nucleotide sequence of the human KLF7 gene promoter is selected from any one sequence as shown in SEQ ID NO: 1 to SEQ ID NO: 18; According to the invention, DNA extracted from human semen is taken as a template, 18 human KLF7 promoters are obtained by using primers for PCR amplification and restriction enzyme digestion, and test verification shows that the 18 promoters all have promoter activity in chicken preadipocytes, so that the human KLF7 gene promoter has important significance for revealing a regulation mechanism of expression of a plurality of human KLF7 transcription spliceosomes and developing drugs for targeting KLF7 expression, and lays a foundation for researching treatment of a plurality of human diseases regulated by the KLF7 gene.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a human KLF7 gene promoter and its construction method and application. Background technique [0002] Krüppel-like factors (Kruppel-like factors, KLFs) are a class of transcription factors present in animals, they are involved in the regulation of multiple life processes such as cell proliferation, differentiation and apoptosis. At present, a total of 17 kinds of KLF factors have been found in the human body. Their protein structure is characterized by three typical C2H2 zinc finger structures at the carboxyl terminus (C-terminus) as the DNA binding domain, and the amino terminus (N-terminus, N-terminus). ) sequence as a transcriptional regulatory domain is poorly conserved among family members. In 1998, Matsumoto et al. cloned a previously unreported KLF factor from human vascular endothelial cells by degenerate PCR technology, and named it the ubiquitous Krüppel-li...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/85C12N5/10C12N15/11A61K45/00
CPCC12N15/11C12N15/113A61K45/00C12N15/85C12N5/10
Inventor 张志威陈月婵李瑶瑶高玲羽张燚吴燕名李泽泉吴湘祁
Owner SHIHEZI UNIVERSITY
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