Atrial fibrillation peccant gene and use thereof

A technology of amino acids and polynucleotides, applied in genetic engineering, plant genetic improvement, application, etc.

Inactive Publication Date: 2006-03-15
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no relationship between KCNE2 and human

Method used

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  • Atrial fibrillation peccant gene and use thereof
  • Atrial fibrillation peccant gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Identification of YQXM

[0104] The strategy of identifying YQXM adopted in the present embodiment is: (1) studies have shown that the mechanism of occurrence of familial atrial fibrillation may be related to ion channels; Exon sequencing screening of candidate potassium channel genes (KCNQ1, HERG, KCNE1, KCNE2, KCNE3, KCNE4, KCNE5, and KCNJ2) to find possible pathogenic gene mutations; (3) Preliminary identification of YQXM mutations in two families After the relationship with atrial fibrillation, further rule out the possibility that more than a certain number of normal random individuals have the same gene mutation; (4) perform mutant function analysis with patch clamp, and finally establish that the YQXM or the encoded protein causes human atrial fibrillation.

[0105] In this embodiment, the steps for determining and identifying YQXM according to the above strategy are: (1) determining the candidate gene; (2) sequencing screening of the candidate gene; (3) excludin...

Embodiment 2

[0125] Establishment of Animal Model of Inherited Atrial Fibrillation

[0126] The human KCNE2 cDNA probe was used to scan the 129Sv mouse genome library, and the mouse genome KCNE2 gene was cloned. After confirmation by PCR, restriction analysis and shotgun sequencing, the fragments located upstream and downstream of KCNE2 in the mouse were selected and joined to the nucleotide sequence containing the nucleotide sequence shown in SEQ ID NO.1 and the neomycin resistance gene and TK gene, Specific homologous recombination at the mouse KCNE2 gene locus. Enabling transfected ES cells to undergo dual drug selection (G418 and Gancyclvir) narrows the scope for screening homologously recombined ES cell clones. The obtained ES cell clones were confirmed by PCR and Southern blot analysis to confirm their correct homologous recombination, and were injected into the blastocysts of C57BL / 6 strain mice after karyotype analysis, and then the injected blastocysts were implanted In utero of...

Embodiment 3

[0129] Preparation and purification of protein encoded by YQXM

[0130] In this example, the full-length YQXM sequence or fragment was constructed into a commercially available protein fusion expression vector, prokaryotically expressed in Escherichia coli in the form of a GST fusion protein, and the recombinant protein was purified.

[0131] (1) Construction of prokaryotic expression vector and transformation of Escherichia coli

[0132] According to the full-length coding sequence of the YQXM encoded protein (SEQ ID NO.1), primers were designed to amplify the complete coding reading frame (corresponding to about 20 or more nucleotides at the 5' and 3' ends of the coding sequence, respectively), and Restriction endonuclease sites were introduced into the anti-primers (this depends on the pGEX-2T vector selected), so as to construct the expression vector. Using the amplified product of YQXM obtained in Example 1 as a template, after PCR amplification, YQXM was cloned into the...

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Abstract

YQXM and its coding protein are both disclosed. YQXM is a mutant of KCNE2 gene and the mutant point happens in No.219 bit of KCNE2. Also, Preparation and use of their nucleotide sequence and coding protein are involved.

Description

technical field [0001] The invention relates to the fields of molecular genetics, cell biology and cardiology. Specifically, the present invention relates to atrial fibrillation pathogenic gene YQXM and its encoded protein. In addition, the present invention also relates to the preparation method and application of YQXM nucleotide sequence and encoded protein. Background technique [0002] According to the definition of the World Health Organization, atrial fibrillation (atrial fibrillation) is irregular and chaotic electrical activity in the atrium. Complete ventricular rate variability in the absence of high-grade or complete AV block. [0003] Atrial fibrillation is one of the most common clinical arrhythmias, which can trigger heart failure or hemodynamic disorders, induce fatal arrhythmias such as ventricular tachycardia or ventricular fibrillation, lead to thrombosis and embolism events, and increase the mortality rate of underlying heart disease , Atrial fibrillati...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K14/47C07K16/18C12N15/12C12N15/63C12P21/02A01K67/00A61K49/00
Inventor 陈义汉刘懿杨奕清
Owner TONGJI UNIV
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