The International Institute for Science, Technology and Education (IISTE)
Abstract
Recombinant DNA domain remains a dependant of effective use of recombinant proteins in many applications but its production remains an area of concern due to DNA vector and production path. This work is aimed at studying the generation and replication of the recombinant DNA molecule using PI3K C2β C2 domain and glutathione s-transferase (GST) which expressed in pGEX-2T vector. The molecular cloning technique was employed to generate a recombinant DNA molecule. PI3K C2β C2 domain of isoform PI3K C2β belonging to class PI3K C2 of Phosphoinositide-3-Kinase (PI3K) family was used and the effective digestion of pGEX-2T vector was studied using restricted enzymes (RE) of EcoR I and Sma I with binding partners location. The ligated product of recombinant DNA was obtained after successful doubled digestion of pGEX-2T vector. The best transformation of competent bacterial cells was obtained at molar ratio of 5:1 for RE digested vector insert DNA and vector DNA. The recombinant DNA can be employed may be used in treatment of non – communicable diseases such as cancer and diabetes. Keywords: Recombinant DNA molecule, Phosphoinositide-3-Kinase (PI3K), PI3K C2β C2 domain, Restriction enzyme, Ligatio
