LIU Zhang-ren, LIU Wei, ZHAI Jin-ling, ZI Liang, HUANG Xi. Construction and Autoactivation Test of a Bait Vector of AtGRP7 for Two-hybrid System[J]. Journal of Tropical Biology, 2012, 3(2): 121-125. doi: 10.15886/j.cnki.rdswxb.2012.02.002
| Citation:
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LIU Zhang-ren, LIU Wei, ZHAI Jin-ling, ZI Liang, HUANG Xi. Construction and Autoactivation Test of a Bait Vector of AtGRP7 for Two-hybrid System[J]. Journal of Tropical Biology, 2012, 3(2): 121-125. doi: 10.15886/j.cnki.rdswxb.2012.02.002
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Construction and Autoactivation Test of a Bait Vector of AtGRP7 for Two-hybrid System
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Key Laboratory of Tropical Biological Resources of Ministry of Education/Institute of Bio-Science and Biotechnology, College of Agronomy, Hainan University, Haikou 570228, China
- Received Date: 2012-02-14
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Abstract
AtGRP7 is one of slave oscillator of Arabidopsis thaliana,but its physiological role is so far less known.In order to screen the AtGRP7 interacted proteins,the CDS sequence of AtGRP7 was first amplified using PCR and inserted into pGBKT7 vector between the clone site NcoⅠ and PstⅠ to construct the bait vector for yeast two-hybrid（Y2H） system.Restricted digestion and sequencing result showed that the AtGRP7 sequence was inserted into pGBKT7 vector in correct reading frame.Furthermore,the bait vector was transformed into Y2H Gold yeast strain to test the autoactivation and toxicity.The transformed yeast grew very well on SD/-Trp plate,but weakly on SD/-Trp/-Ade plate.No yeasts grew on SD/-Trp/-His and SD/-Trp/-Ade/-His plates,suggesting that histidine synthase gene was not transcribed and translated.This indicated that AtGRP7 was not autoactivated in the Y2H system.
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