BRCA2 gene mutant and application thereof

A mutant and coding technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as chromosomal instability, loss of protein to repair DNA damage, etc., and achieve the effect of improving and enriching the diagnostic process.

Inactive Publication Date: 2018-03-06
SHENZHEN HUADA GENE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein expressed after the BRCA2 gene mutation loses the function of repair

Method used

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  • BRCA2 gene mutant and application thereof
  • BRCA2 gene mutant and application thereof
  • BRCA2 gene mutant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Determination of BRCA2 gene mutation in hereditary breast cancer

[0036] 1. Experimental materials

[0037] Sample information: a family with hereditary breast cancer, including a proband and 5 family members, see Table 2 for details

[0038] Main instrument information: pipette, PCR instrument, centrifuge, vortex shaker, electrophoresis instrument, DNA breaker, magnetic stand, dry constant temperature metal bath, Complete Genomics sequencer

[0039] Main reagents: QIAGEN Blood DNA Extraction Kit, Complete Genomics platform library building reagents, liquid phase probe of about 600Kb target region in the human genome (hg19) (Roche, Nimblegen EZ chip)

[0040] 2. Sample collection

[0041] Such as figure 2 As shown, the hereditary breast cancer family contains 6 members, including 4 breast cancer patients. A breast cancer patient was selected as the proband in the family, and the regional capture and sequencing samples were taken. The basic information ...

Embodiment 2

[0064] Example 2 Verifying the causative mutation site using the gold standard (Sanger sequencing)

[0065] In order to verify the technical accuracy of the invention, PCR primers were designed for the region where the mutation site is located, and the primer sequences are shown in Table 4. After conventional PCR amplification, Sanger sequencing was performed to analyze the peak pattern of the target site.

[0066] Table 4. BRCA2 gene c.3467delC mutation site PCR primer sequence

[0067]

[0068] PCR reaction system: 40μL

[0069]

[0070] PCR reaction conditions

[0071]

[0072] The verification results show that the analysis results of the high-throughput sequencing CG sequencing platform are consistent with the results of Sanger sequencing, and the peak image obtained by Sanger sequencing is as follows figure 1 shown.

Embodiment 3

[0073] Example 3 Carrying out mutation verification and family history analysis on relative family members of mutation carriers

[0074] Using PCR combined with Sanger sequencing method, the family members of the proband with a breast cancer susceptibility gene mutation were verified for the corresponding mutation site to determine whether the mutation site was co-segregated with the disease. The family diagram is as follows figure 2 shown.

[0075] The verification process is as follows:

[0076] (1) DNA extraction

[0077] Genomic DNA was extracted from blood samples of breast cancer patients using the QIAGEN DNA Blood mini kit and following the kit’s extraction instructions. Detect the DNA concentration. In principle, the amount of DNA obtained from each sample is ≥ 2 μg, and then electrophoresis is used to detect whether the DNA is complete and its degree of degradation.

[0078] Design PCR primers for the region where the mutation site is located, perform Sanger sequ...

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PUM

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Abstract

The invention discloses a BRCA2 (breast cancer) gene mutant and application thereof, and particularly relates to a separated nucleic acid of an encoded BRCA2 mutant, a separated polypeptide, a methodfor detecting a BRCA2 gene mutation site, and a system and a kit for detecting the BRCA2 gene mutation site, wherein the separated nucleic acid of the encoded BRCA2 mutant has c.3467delC mutation whenbeing compared with SEQ ID NO:1. By detection whether a new mutant exists in a biological sample or not, the BRCA2 gene mutant can effectively detect whether the biological sample is liable to get the hereditary breast cancer.

Description

technical field [0001] The present invention relates to BRCA2 gene mutant and application thereof. Specifically, the present invention relates to isolated nucleic acids encoding BRCA2 mutants, isolated polypeptides, and methods, systems and kits for detecting BRCA2 gene mutants. Background technique [0002] In recent years, the morbidity and mortality of cancer in my country have been increasing, and malignant tumors have become a major public health problem in China. According to the results released by the my country Cancer Registry Center in 2016, in 2012, there were 3.586 million new cancer cases in my country, 2.187 million deaths, a crude incidence rate of 265 / 100,000, and a mortality rate of 161.5 / 100,000. In the current situation of malignant tumors, breast cancer ranks first among female malignant tumors, with about 273,000 new cases every year, and the age of onset is showing a trend of younger age. It is the number one killer that seriously threatens women's liv...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12Q1/6886C07K14/47G01N33/68C12M1/34
CPCC07K14/4703C12Q1/6886C12Q2600/156G01N33/68G01N2333/4704
Inventor 刘梦王晓宏邵康曹博洋李贵波侯勇朱师达
Owner SHENZHEN HUADA GENE INST
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