Pura gene segment P2 and application thereof

A technology of α gene and fragment, applied in Purα gene fragment P2 and its application field, can solve the problem of rare Purα mutant

Inactive Publication Date: 2016-11-16
NINGXIA MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] At present, in the prior art, there are few studies on the construction of Purα mutant expressed in fusion with GFP protein

Method used

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  • Pura gene segment P2 and application thereof
  • Pura gene segment P2 and application thereof
  • Pura gene segment P2 and application thereof

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Experimental program
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Embodiment Construction

[0081] The technical solutions of the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.

[0082] A Purα gene fragment P2, the nucleotide sequence of which is shown in SEQ ID NO:1.

[0083] The application of the Purα gene fragment P2 of the present invention in the process of constructing a Purα mutant expressed by fusion with GFP protein.

[0084] Further, the application of the Purα gene fragment P2 in the process of constructing a Purα mutant fused and expressed with GFP protein includes the following steps:

[0085] 1.1 Design of primers

[0086] The protein structure of Purα consists of three major parts: N-terminal glycine-rich region; C-terminal psychomotif and glutamine- and glutamate-rich regions and a central DNA-binding region. Its DNA-binding region contains 3 class I and 2 class II repeats, located in the amino acid regions 66-88, 102-131, 148-170, 188-120 and 224-246, respectively, ...

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Abstract

The invention discloses a Pura gene segment P2 and an application thereof. The nucleotide sequence of the gene segment P2 is shown as SEQ ID NO:1. According to the Pura gene segment P2 disclosed by the invention, the Pura gene segment on a pCDNA3.0 vector is subjected to eukaryotic expression, a plasmid is transfected into a required research cell line and Western immunoblotting is conducted so as to discuss the effects of different structural domains of Pura protein on a target gene. The Pura gene segment on the pEGFP-C1 vector can achieve fusion expression with green fluorescent protein GFP, the distribution of the Pura protein in cells is observed by virtue of a fluorescence microscope, and the expression levels of the Pura proteins in the various structural domains are quantified on the basis of the expression level of the GFP protein. By virtue of the Pura protein gene segment on a prokaryotic expression vector pGEX, the effects of the different structural domains of the Pura protein on the target gene can be researched in vitro.

Description

technical field [0001] The invention belongs to the field of biotechnology, and discloses a Purα gene fragment P2 and an application thereof. Background technique [0002] Purα has a motif structure, and its central part is the DNA binding region, which contains three first-order repeats and two second-order repeats. Other notable structural features include an N-terminal glycine-rich region, a "Psycho" motif region and C-terminal glutamine and glutamate-rich regions. The sequence of the DNA-binding region of Purα is extremely conserved throughout evolution. Purα is expressed in almost all metazoan tissues. It is a multifunctional protein that can bind to both DNA and RNA. It plays a role in the initiation of DNA replication, mRNA transcription and protein translation to varying degrees. effect. In neurons, Purα can be transported and bound to mRNA. Purα is closely related to the DNA sequence of the origin of replication of viruses and cells. Since the initiation of repl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K19/00C12N15/70
CPCC07K14/47C07K2319/60C07K2319/80C12N15/70C12N2800/101
Inventor 崔建奇李永玲贾中发
Owner NINGXIA MEDICAL UNIV
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