Revista Europea de Biología Experimental Acceso abierto

Abstracto

Genotyping of Her1 SNP’s in familial breast cancer by restriction fragment length polymorphism and sequencing

Ibtihal Riyadh Najeeb, Sudhakar Malla, Radhakrishnan Senthilkumar and Selvam Arjunan

Although the incidence of many types of cancer has declined globally over the last thirty years, the incidence of breast cancer globally has increased. Among women in worldwide, breast cancer remains one of the most common cancers. Genetic changes can occur at different levels and by different mechanisms. The gain or loss of an entire chromosome can occur through errors in mitosis. More common are mutations, which are changes in the nucleotide sequence of genomic DNA. Novel methods are being developed for the production of antibodies to specific antigens and thus helping in the process of development of protein based vaccines. HER1 gene was amplified from the tissue samples of the infected persons. In our present study 15 Indian families with hereditary breast cancer were studied for HER1 mutations using PCR RFLP method. Blood sample were collected and total genomic DNA was isolated using modified CTAB method. The isolated DNA was run on 1% agarose gel to verify the quality and the quantity was estimated by spectophotometrically. PCR was done using the specific primer and the gene product was electrophorized in 2% agarose gel. The amplified product was subjected to restriction digestion using EcoR1 and the PCR RFLP pattern was analyzed. The sample no 11 showed a single band, which is not digested with the restriction enzyme. So it confirmed the mutation in the recognition site of the enzyme. We eluted the PCR product along with two normal sample and send for sequencing. The sequencing result showed that there is a single mutation at the EcoR1 restriction site. So RFLP can be used as to find out mutation or SNP in the gene responsible for the cancer development.