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
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.
