TRANSFORMATION OF GENE TcPIN THE EXPRESSION VECTOR pBIN+ FOR SOYBEAN POD BORER RESISTANCE
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Date
2014-05-22
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Abstract
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
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Keywords
TcPIN gene, soybean pod borer, gene constuction, proteinase inhibitor