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异戊烯基转移酶基因作为烟草(烟草品种 Petit Havana SRI)植物转化的选择标记基因是有效的。

The isopentenyl transferase gene is effective as a selectable marker gene for plant transformation in tobacco (Nicotiana tabacum cv. Petite Havana SRI).

作者信息

Endo S, Kasahara T, Sugita K, Matsunaga E, Ebinuma H

机构信息

Pulp and Paper Research, Nippon Paper Industries, Oji 5-21-1, Kita-ku, Tokyo 114-0002, Japan e-mail:

出版信息

Plant Cell Rep. 2001 Jan;20(1):60-66. doi: 10.1007/s002990000279.

Abstract

A selection method for transformed cells which does not inhibit regeneration is important for the establishment and optimization of a transformation protocol. We have assessed the 35S-ipt gene from Agrobacterium tumefaciens as a selectable marker gene. The identification of ipt-expressing cells from nontransformed cells enabled morphological selection without the use of kanamycin and also allowed for the elimination of a high proportion of nonexpressing cells. Ipt selection of tobacco leaf discs (Nicotiana tabacum cv. Petite Havana SRI) resulted in a 2.7-fold higher transformation frequency compared to kanamycin selection. Overexpression of the ipt gene favored plant regeneration from transformed cells, and the transformation frequency of the ipt plus kanamycin selection resulted in a 1.6-fold higher transformation frequency than kanamycin selection alone. These results indicate that this procedure might provide a strategy whereby transgenic plants can be efficiently obtained and some of the problems related to the use of antibiotics diminished.

摘要

一种不抑制再生的转化细胞筛选方法对于转化方案的建立和优化至关重要。我们评估了来自根癌农杆菌的35S-ipt基因作为选择标记基因。从未转化细胞中鉴定出表达ipt的细胞能够在不使用卡那霉素的情况下进行形态学筛选,并且还能消除很大比例的不表达细胞。对烟草叶盘(烟草品种Petite Havana SRI)进行ipt筛选,与卡那霉素筛选相比,转化频率提高了2.7倍。ipt基因的过表达有利于转化细胞的植株再生,ipt加卡那霉素筛选的转化频率比单独使用卡那霉素筛选高出1.6倍。这些结果表明,该方法可能提供一种策略,据此可以高效获得转基因植物,并减少与使用抗生素相关的一些问题。

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