%A QI Jing, DONG Zhen, ZHANG Yu-Xing %T Cloning of ACC Oxidase Gene from Yali Pear and Tranformation of Its Antisense Expression Vector with Agrobacteriummediated Method %0 Journal Article %D 2014 %J Plant Diversity %R 10.7677/ynzwyj201413225 %P 622-628 %V 36 %N 05 %U {https://journal.kib.ac.cn/CN/abstract/article_33355.shtml} %8 2014-09-25 %X

In this study, a partial ACC oxidase (ACO) genelike cDNA sequence was obtained through homologybased cloning from Yali (Pyrus bretschneideri cv. ‘Yali’) plant. Primers were designed according to the highly conserved regions of published ACO gene sequences, and RTPCR cloning was conducted by using Yali fruit cDNA. The obtained ACOlike cDNA fragment contains 831 base pairs which encodes 276 predicted amino acid residues, and shares no less than 94% nucleotide sequence identity with all published ACO genes. We further inversely inser ted the ACOlike cDNA fragment into pBI121 expression vector, and transformed it into tissue cultured Yali plants by using Agrobacterium LBA4404. Finally, 4 independent transgenic lines harboring the antisense ACOlike fragment were obtained and validated by PCR analysis. Southern blotting assay revealed 3 transgenic lines containing single copy of the foreign gene, and 1 line with 2 insertion copies.