%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 Agrobacteriummediated 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) genelike cDNA sequence was obtained through homologybased cloning from Yali (Pyrus bretschneideri cv. ‘Yali’) plant. Primers were designed according to the highly conserved regions of published ACO gene sequences, and RTPCR cloning was conducted by using Yali fruit cDNA. The obtained ACOlike 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 ACOlike 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 antisense ACOlike 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.