Abstract

Khairy MA Zoheir , and Li-Guo Yang

Gene cloning, or molecular cloning, has several different meanings to a molecular biologist. A clone is an exact copy, or replica, of something. Molecular biologists exploit the replicative ability of cultured cells to clone genes. Recent advances in the Restriction enzymes technologies have opened up new opportunities in biotechnology and facilitate the way to isolate any gene from any source of cells. Many types of restriction enzymes cleave DNA away from their recognition site. By using the type III restriction enzyme, we provide evidence to show that an intact recognition site on the cleaved DNA sequesters the restriction. We isolated and cloned the IGF-1 R gene from DNA by subjecting DNA with one pair of restriction enzyme (EcoRI + HindIII) and followed by ligation and transformation with the suitable vector. The pair of restriction enzymes (EcoRI + HindIII) which subjected to DNA template before the ligation process was the only pair which had success to liberate IGF-1R fragment from the transformed vector rather than other two pairs of restriction enzymes (EcoRI + XhoI) or (HindIII + XhoI) although this two pair of restriction enzymes have recognition sites in the vector. Moreover we have studied the effect of DNA template size on the action of the restriction enzymes. Where less DNA size, more efficient of the restriction enzyme action. This study is rapid and useful new technology for isolation, cloning and sequencing the gene of interest to be ready for further investigations as gene transfection and/ or transfer