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Puralpha gene segment P4 and application thereof

A technology of α gene and fragment, which is applied in the Purα gene fragment P4 and its application field, and can solve the problems such as the rareness of Purα mutants

Inactive Publication Date: 2016-11-16
NINGXIA MEDICAL UNIV
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  • Abstract
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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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  • Puralpha gene segment P4 and application thereof
  • Puralpha gene segment P4 and application thereof
  • Puralpha gene segment P4 and application thereof

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Embodiment Construction

[0076] The technical solution of the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0077] A Purα gene fragment P4, the nucleotide sequence of which is shown in SEQ ID NO:1.

[0078] The application of the Purα gene fragment P4 of the present invention in the process of constructing the Purα mutant expressed in fusion with GFP protein.

[0079] Further, the application of the Purα gene fragment P4 in the process of constructing the Purα mutant expressed in fusion with GFP protein comprises the following steps:

[0080] 1.1 Design of primers

[0081] The protein structure of Purα includes three parts: the N-terminal glycine-rich region; the C-terminal psycho motif and glutamine- and glutamate-rich region and the central DNA-binding region. Its DNA-binding region contains 3 class I and 2 class II repeats, located in the 66-88, 102-131, 148-170, 188-120 and 224-246 amino acid regions, such as...

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Abstract

The invention discloses a Puralpha gene segment P4 and application thereof. A nucleotide sequence of the Puralpha gene segment P4 is shown as SEQIDNO:1, the Puralpha gene segment on a pCDNA3.0 vector undergoes eukaryotic expression, the plasmid is transfected to a required study cell line, and the effects on target genes of different structural domains of Puralpha proteins are explored through a protein immunoblot experiment. The Puralpha gene segment on a pEGFP-C1 vector and a green fluorescent protein (GFP) undergo fusion expression, the distribution situation of the Puralpha proteins in cells is observed under a fluorescence microscope, and the expression levels of the different structural domains of the Puralpha proteins in the cells are quantified through the expression level of the GFP protein. In-vitro study can be conducted on the effects on the target genes of the different structural domains of the Puralpha proteins through the eukaryotic expression conducted on the Puralpha protein gene segment on a pGEX vector.

Description

technical field [0001] The invention belongs to the field of biotechnology and discloses a Purα gene fragment P4 and its application. Background technique [0002] Purα has a motif structure, and its central part is a DNA-binding region, which contains three 1st-level repeats and two 2nd-level repeats. Other notable structural features include the N-terminal glycine-rich region, a "Psycho" motif region and the C-terminal glutamine and glutamic acid-rich regions. The sequence of the DNA-binding domain 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 DNA replication initiation, mRNA transcription and protein translation to varying degrees. effect. In neurons, Purα can be transported and bound to mRNA. Purα has a close relationship with the DNA sequence of the replication initiation site of viruses and cells. Since the initiation of rep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/70C12N15/65
CPCC07K14/435C12N15/65C12N15/70
Inventor 崔建奇李永玲贾中发
Owner NINGXIA MEDICAL UNIV
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