PJB-2012-311
CONSTRUCTION OF EUKARYOTIC PLANT EXPRESSION VECTOR WITH THE SAG12 PROMOTER AND THE LEAF SENESCENCE–RELATED IPT GENE AND ITS GENETIC TRANSFORMATION IN RICE
PINGZHONG CAI1,2, QIUFANG QIAN1, ZHENHUI KANG1, ZHIYONG ZHANG2 AND GUIXUE WANG1*
Abstract
A pair of primers was designed according to the sequences published by GenBank to amplify the isopentenyl transferase gene (ipt gene) of the plasmid in Agrobacterium tumefaciens (strain C58). The 5′-untranslated sequence of the senescence-associated SAG12 gene of Arabidopsis was isolated by the polymerase chain reaction (PCR). In addition, we constructed the plant-expression vectors (pBI121-ipt and pCAMBIA1301-SAG12-ipt), which were regulated by the SAG12 promoter and the ipt gene. The plant-expression vector was used to transform the embryos of rice cultivar Zhonghua 16 with the help of the A. tumefaciens system. Plantlets were regenerated in vitro by selection on medium containing hygromycin. PCR amplification showed that the target gene was successfully integrated into the transformed plants.
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