Team:KAIST-Korea/Project/Methods
From 2010.igem.org
PCR
To manipulate gene products obtained from gene-bank, our team used PCR/Real-Time PCR methods. Commercially obtained genes were containing few more base paires added at front and back site. As FGPR, the receptor protein of human, has human-specific signal peptide at its starting point, this region had to be replaced
by S.pombe specific signal peptide to express the protein at membrane region. To do this, PCR primer containing signal peptide region of S.pombe were synthesized with method described below. Then, FGPR gene was amplifed by PCR to get gene product which its signal region is replaced. Also, additional sequence at rear site of gene was
removed. STAT gene passed through same process, removing additional region and adding required signal peptide.
PCR was done by commercial service sponsered by Bioneer.Inc (Korea), using AccuPower PCR system supplied by same company as reference below.