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The peaT1 gene fragment was amplified from pGEM-6p-1-peaT1 by PCR,and recovered target gene was cloned into pLexA vector.After digestion and sequencing,the bait vector pLexA-peaT1 was transformed into yeast strain EGY48 [p8op-lacZ] by PEG/LiAC,and the transcriptional activity of bait vector was detected.The results showed that recombinant bait plasmid pLexA-PEMG1 was constructed,for the two bands of recombinant bait plasmid in agarose gel electrophoresis were expected after digesting by restriction endonuclease EcoR I and Xho I.Therefore,the recombinant bait plasmid could be used in yeast two-hybrid system to screen a cDNA library.
The peaT1 gene fragment was amplified from pGEM-6p-1-peaT1 by PCR, and the recovered target gene was cloned into pLexA vector. After digestion and sequencing, the bait vector pLexA-peaT1 was transformed into yeast strain EGY48 [p8op- lacZ] by PEG / LiAC, and the transcriptional activity of bait vector was detected. The results showed that recombinant bait plasmid pLexA-PEMG1 was constructed, for the two bands of recombinant bait plasmid in agarose gel electrophoresis were expected after digesting by restriction endonuclease EcoR I and Xho I. Therefore, the recombinant bait plasmid could be used in yeast two-hybrid system to screen a cDNA library.