Breast cancer related protein, gene encoding the same, and method of diagnosing breast cancer using the protein and gene
a breast cancer and related protein technology, applied in the field of breast cancer related protein, can solve the problems of increasing the chance of developing breast cancer, overexpression of an oncogene, and considerably slowing the progress of breast cancer
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example 1
Search of BCRP Gene Based on Single Nucleotide Polymorphism (SNP) Data
[0066] 1. Search of BCRP Gene
[0067] The nucleotide sequence of the site, at which the SNP was found was searched through a database (NCBI) search and analysis. The association of the SNP with breast cancer was accomplished in a separate study. The information suitable for fabricating a primer for amplifying a gene was obtained therefrom.
[0068] 2. Amplification of BCRP Gene Fragment
[0069] A primer for amplifying DNA around the searched SNP was designed using the sequence information obtained through the database (SEQ ID Nos. 1 and 2).
[0070] Next, PCR was performed using genomic DNA as a template and the designed primer set as a primer so that BCRP gene fragment of 239 bp was amplified. In the PCR, 10 pmol of each of forward and reverse primers (SEQ ID Nos. 1 and 2) and 200 pg-1 μg of genomic DNA template were used and the reaction was 35 times repeated at 95° C. for 40 seconds, at 57° C. for 40 seconds, and 72...
example 2
Identification of Expression of BCRP Gene in Cells and Tissues
[0077] 1. Identification of Expression of BCRP Gene in Cells through Northern Blotting
[0078] A northern blotting analysis was performed on a plurality of human normal tissues and tumor tissues (available from Clontech Corp.) using the PCR product obtained from Example 1 as a probe. The results are illustrated in FIGS. 1 and 2. Referring to FIGS. 1 and 2, the BCRP gene was specifically expressed only in heart tissue among normal tissues and in breast cancer tissue among tumor tissues. Consequently, it is apparent that the expression of the BCRP gene can be used to detect the presence of breast cancer. In FIG. 1, tissues used in the northern blotting were brain heart, heart, skeletal muscle, colon, thymus, spleen, kidney, liver, small intestine, placenta, lung, and peripheral blood leukocyte cell. In FIG. 2, cancer tissues used in the northern blotting were cell lines of breast, ovary, uterine, lung, kidney, stomach, colo...
example 3
Identification of Location of Expression of BCRP Gene in Cells
[0095] The localization of the expression of the BCRP gene in cells was identified using an immunocytochemistry method. For this experiment, the BCRP gene was first cloned to pFLAG vector (Sigma, Amherst, N.Y.) using recombinant DNA techniques. The cloning procedure was as follows. The BCRP gene of SEQ ID No. 3 was digested with Not I enzyme and Sal I enzyme, the pFLAG vector was digested with the same enzymes, and then the digested BCRP gene and the pFLAG vector were ligated. The cloned BCRP-pFLAG vector DNA was transfected to various cell lines using Lipofection 2000. The transfected cells were incubated in 5% CO2 at 37° C. for 48 hours such that the BCRP gene could be expressed. Then, the incubated cells were fixed on a plate using 3.5% paraformaldehyde. To dye the inside of the cell, the cell was changed to be permeable by 0.1% Triton X-100. A blank space was blocked with 1% BSA blocking solution. A flag specific ant...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Fraction | aaaaa | aaaaa |
| Level | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More