Investigation of inducible transposon to remove selection marker

碩士 === 國立臺灣大學 === 農藝學研究所 === 93 === The maize transposable element Ac can move to new location within genome. In this work, an Ac–based inducible transposon was constructed to develop a one-time gene expression system. Firstly, the 5’ end of the Ac was inserted in the first intron of the modified ri...

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Bibliographic Details
Main Authors: Kuan-Te Li, 李冠德
Other Authors: Yuh-Chyang Charng
Format: Others
Language:zh-TW
Published: 2005
Online Access:http://ndltd.ncl.edu.tw/handle/89998326962996173504
Description
Summary:碩士 === 國立臺灣大學 === 農藝學研究所 === 93 === The maize transposable element Ac can move to new location within genome. In this work, an Ac–based inducible transposon was constructed to develop a one-time gene expression system. Firstly, the 5’ end of the Ac was inserted in the first intron of the modified rice EPSPS gene, which triggered by the CaMV35S promoter. The inducible transposon was completed by locating the PR-1a promoter-transposase fusion and the CaMV35S promoter was between the 5’ and 3’ ends. Thus, this inducible transposon contains not only the PR-TPase fusion but also the first exon of the modified EPSPS gene and its promoter. This construct, which termed as KCEH, was introduced into rice. The modified EPSPS gene can be transcribed normally and allow the transgenic rice to be Glyphosate-resistant. After the Glyphosate screening, the transgenic rice plants were treated with salicylic acid for the excision of the inducible transposon. As a result of this, the modified EPSPS gene was truncated by losing its first exon and the 35S promoter. Emphasis has been placed on the theoretical and practical aspects of the approach that may be relevant to its application to other gene expression regulatory system. It provides a means to investigate a one-time gene expression system.