You have detected and purified a novel protein that highly e

You have detected and purified a novel protein that highly expressed in cancerous liver cells in chickens. You name the Protein XFILE in honor of some TV show you used to watch. This protein is found in the cytoplasm and is apparently 25 kDa in mass. 1) Edman degradation gives you the following amino acids data from the N-terminus

Solution

The nucleotide sequence to the protien shown is

a) Both the primers in 5\' -3\' direction

Forward Primer ATGAGCGATGTGGCCATC

Reverse primer GGGCAGCAGCTCGAACTT

b) The genomic DNA from chicken will be isolated by DNA extraction kit using manufactures protocol. The DNA will be purified. Primers will be designed for the specific gene of interest. The gene is then amplified by Polymerase chain reaction(PCR). The PCR product is analysed by agarose gel electrophorosis and then purified from the gel. The PCR product is then sequenced by sanger sequencing.

c) A flouroscent tagged antibody will be used against the protien of interest. The cells will be stained by DAPI which will stain all the cells. The protien will be stained and the presence can be located by the flouroscent staining using a flourosscence microscopy.

2a) The same gene is searched in NCBI nucleotide sequence/

b) Human DNA is isolated from human blood sample, DNA is purified and then using specific primers that will amplify the gene of interest the gene is amplified. The PCR product is purified and sequenced.