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Primer set, kit and detection method for detecting mutations in the tnf coding region of exoprotein a gene

A technology of foreign protein and coding region, which is applied to the primer set for detection of mutation of TNF coding region of foreign protein A gene, and the application field of the kit to achieve the effects of accurate results, simple detection process and wide detection range.

Inactive Publication Date: 2021-03-26
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no method for detecting mutations in the TNF coding region of the foreign protein A gene in population samples and judging the phenotype based on the gene mutation. Therefore, establishing a method for detecting mutations in the TNF coding region of the foreign protein A gene and detecting and predicting mutations may lead to The phenotype is imminent

Method used

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  • Primer set, kit and detection method for detecting mutations in the tnf coding region of exoprotein a gene
  • Primer set, kit and detection method for detecting mutations in the tnf coding region of exoprotein a gene
  • Primer set, kit and detection method for detecting mutations in the tnf coding region of exoprotein a gene

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Experimental program
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Effect test

Embodiment 1

[0055] Using the human peripheral blood genomic DNA of the sample a to be tested as a template, three pairs of primers from exons 7, 8, and 9 were used to perform PCR reactions, and the amplified products were sequenced, and the sequence comparison software MEGA7 was used to compare the sequencing results with wild-type EDA Align the sequences of coding regions of exons 7, 8, and 9 of genes with SEQ ID No.7, SEQ ID No.8, and SEQ ID No.9 respectively, or splice parts of exons 7, 8, and 9 in the sequencing results, and The wild-type EDA gene TNF coding region sequence SEQ ID No.10 is compared to determine whether there is a mutation and the specific mutation site.

[0056] Further, the sequence 1, sequence 2, and sequence 3 of the above sequencing results were spliced ​​to obtain the continuous sequence of exon 7, 8, and 9 of the sample to be tested, and translated into an amino acid sequence, which was compatible with the EDA protein TNF The domain amino acid sequence SEQ ID No...

Embodiment 2

[0077] Using the human cultured cell genomic DNA of the sample b to be tested as a template, design 3 pairs of primers targeting its exons 7, 8, and 9, perform sequencing after PCR reaction, and use sequence comparison software to compare the sequencing results with the wild-type EDA gene 7, 8, and 9 exon coding region sequences SEQ ID No.7, SEQ ID No.8, and SEQ ID No.9 were compared respectively, or exons 7, 8, and 9 in the sequencing results were partially spliced, and wild Type EDA gene TNF coding region sequence SEQ ID No.10 is compared to determine whether there is a mutation and the mutation site.

[0078] Further, the sequence 1, sequence 2, and sequence 3 of the above sequencing results were spliced ​​to obtain the continuous sequence of exon 7, 8, and 9 of the sample to be tested, and translated into an amino acid sequence, which was compatible with the EDA protein TNF The domain amino acid sequence SEQ ID No.11 is compared to determine whether the TNF coding region o...

Embodiment 3

[0088] Using the human peripheral blood genomic DNA of the sample c to be tested as a template, design 3 pairs of primers targeting exons 7, 8, and 9, perform sequencing after PCR reaction, and use sequence comparison software to compare the sequencing results with the wild-type EDA gene TNF domain 7, 8, 9 exon coding region sequence SEQ ID No.7, SEQ ID No.8, SEQ ID No.9 were compared respectively, or exon 7, 8, 9 in the sequencing results were partially spliced , compared with the wild-type EDA gene TNF coding region sequence SEQ ID No.10 to determine whether there is a mutation and the mutation site.

[0089] Further, the sequence 1, sequence 2, and sequence 3 of the above sequencing results were spliced ​​to obtain the continuous sequence of exon 7, 8, and 9 of the sample to be tested, and translated into an amino acid sequence, which was compatible with the EDA protein TNF The domain amino acid sequence SEQ ID No.11 is compared to determine whether the TNF coding region of...

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Abstract

The invention provides a specific primer group capable of detecting ectodysplasin A gene TNF code region nucleotide sequence, and a kit capable of detecting ectodysplasin A gene TNF code region nucleotide mutation which contains the specific primer group, a detection method thereof and application of the specific primer group and the kit to detection of the ectodysplasin A gene TNF code region nucleotide mutation. The primer group provided by the invention can amplify a single band without non-specificity band under an appropriate condition; the detection process is simple and convenient, only after PCR amplification and sequencing need to be done, the detection specificity is strong, the sensitivity is high and the result is accurate.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a primer set, a kit, and an application of the kit for detecting mutations in the TNF coding region of the foreign protein A gene. Background technique [0002] Congenital dental hypogenesis is an inherited disorder of defective tooth development, usually during tooth germ formation, where permanent teeth fail to develop and form. It is divided into non-syndromic hypodontia (NSH, OMIM No. 313500) and syndromic hypodontia (SH, OMIM No. 305100). Teeth are accompanied by sparse hair and few sweat glands. Congenital dental hypoplasia will seriously affect the patient's chewing, pronunciation, facial appearance, physical and mental development and mental health. [0003] The mutation of ectodysplasin-A (EDA) gene is the decisive factor leading to non-syndromic edentulousness and syndromic edentulousness. The EDA gene (NM_001399) is located on the long arm of the X chromo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 苏莉翁俊胡巧丽杨艳
Owner HUAZHONG UNIV OF SCI & TECH