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Construction and Transformation of Vernalization-related Genes VIN3 Antisense Plant Expression Vector from Cabbage |
YU Xi-hong,ZHAO Rong-qiu,JIANG Xin-mei*,LI Yan-ru |
College of Horticulture,Northeast Agricultural University,Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture,Harbin 150030,Heilongjiang,China |
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Abstract Taking plant expression vector pBI121 and BoVIN3-1 gene segment as bases,constructed vernalization-related gens VIN3 antisense plant expression vector from cabbage(Brassica oleracea var. captata L.),then inserted the BoVIN3-1 gene segment into XbaⅠand SmaⅠrestriction enzymes site between CaMV355 promoter and GUS gene in a reverted direction,and constructed plasmid pBI35S-BoVIN3-1.Through transferring it to cabbage flower bud by microinjection method,9 cabbages with Kan-resistance were obtained.The result of PCR amplifications proved that the obtained 5 cabbage were positive plant,and the positive plant rate was 55.6%.Compared the transgenic plant after vernalization treatment with the contrast,the transgenic plant vernalization response be delayed at certain extent.The result of RT-PCR amplifications proved that there was a little expression of VIN3 at small degree after low temperature treatment,and its transcription level increased with the prolongation of treatment time.The transcription reaches the peak at 50 days of vernalization.
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Received: 15 January 2012
Published: 06 August 2012
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