4.Seminar UR-UKM KE-7 2012

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TRANSFORMATION OF GENE TcPIN THE EXPRESSION VECTOR pBIN+ FOR SOYBEAN POD BORER RESISTANCE
(2014-05-22) Isda, Mayta Novaliza; Chaidamsari, Tetty; Lestari, Wahyu
Potential of gene resistance found in cocoa like proteinase inhibitor (pin) has an important role in plant defense system against predators and pathogens. Utilization of the pin is a new alternative to soybean in pod borer resistance. Proteinase Inhibitor (PIN) is known as a part of the plants defences system towards the pest. The research is conduct to construct TcPIN gene from cocoa crop into expression pBIN+ vector. The first step of the research is designed a new primary for the TcPIN gene construction. The TcPIN gene yielded from the purification of PCR product is construct into a flap plasmid completed with promotor and terminator. And as the next step, the gene fragment is constructed into pBIN+ and transformed inside competence cell of XL-1 Blue to multiply the plasmid. The result of the amplification is digested with the restrict enzyme SalI and BamHI to exposed the insert (PIN gene) from pGEMT-easy vector. The exposed insert (PIN gene) is transform by pFlap before transferred into the expression vector which is pBIN+. The result showed that the gene TcPIN has been inserted into pBIN+. The construction of the recombinant DNA shown with the digestion result of pBIN+, after cutting with Asc1 and Pac1 restrict enzyme