Expression of Arabidopsis multiple genes in transgenic tobacco plants

• Main Author: 羅慧娟
• Other Authors: 張岳隆
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/08423050008018236063

碩士 === 國立嘉義大學 === 生物農業科技學系研究所 === 103 === High-molecular-weight DNA transformation will become more important in the future due to the availability of more genomic sequences of crop plants and long artificial synthetic DNA techniques. Recently, some vectors have been developed for the purpose of the transformation of large DNA fragments into plants. These vectors, called binary bacterial artificial chromosomes (BIBACs), are designed both for large DNA cloning and for Agrobacterium-mediated or bombardment transformation. In this study, the transgenic plants of tobacco were previously obtained from particle bombardment on tobacco leaf tissues with 83 kb BiBAC DNA of Arabidopsis. The transgenic plants have been propagated to T4 generation through the self-pollination procedure. Based on the Arabidopsis genome sequence data at The Arabidopsis Information Resource (TAIR), the 83-kb DNA fragments host around 20 intact genes. For investigating the expression level of each gene in different tissues or organs of haploid and diploid tobacco transgenic plants, reverse transcription polymerase chain reaction (RT-PCR) is used. The genes with high expression level in diploid plants are: AT1G72250.2, AT1G72330.3, AT1G72370.1 and AT1G72416.3 in mature leaves; AT1G72370.1 and AT1G72450.1 in young leaves; AT1G72250.2, AT1G72290.1, AT1G72410.2, AT1G72430.1 and AT1G72450.1 in mature flowers; AT1G72250.2 and AT1G72450.1 in flower buds. The genes with high expression level in haploid plants are: AT1G72250.2, AT1G72330.3 and AT1G72430.1 in mature leaves; AT1G72320.1, AT1G72440.1 and AT1G72450.1 in young leaves; AT1G72290 .1 and AT1G72330.3 in mature flowers; AT1G72370.1 in flower buds.