Kilby N J, Fyvie M J, Sessions R A, Davies G J, Murray J A
Institute of Biotechnology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QT, UK.
J Exp Bot. 2000 May;51(346):853-63.
Stably transformed Arabidopsis lines in which GUS marked cell clones are readily produced in response to heat-shock have been established and characterized. Control of GUS activation is achieved by heat-shock-induced FLP recombinase activity which "switches on" expression of a GUS marker gene previously held transcriptionally silent. To obtain efficient GUS sectoring, single insert Arabidopsis lines carrying FLP recombinase under the control of a heat-shock-inducible promoter and an FLP-activatable GUS construct were generated. Analysis of GUS sectoring in lines hemizygous and homozygous for both inserts was conducted after various regimes of heat-shock were given at various developmental stages. It is shown that GUS sectoring events can be efficiently induced in most vegetative, aerial and sexual structures in Arabidopsis. Furthermore, the frequency of sectoring events, sector size and, to some extent, the tissues in which sectors are generated can be readily controlled by choice of the conditions and timing of heat-shock used.
已经建立并表征了稳定转化的拟南芥株系，在这些株系中，GUS标记的细胞克隆能够在热激响应下轻松产生。通过热激诱导的FLP重组酶活性实现对GUS激活的控制，该活性“开启”先前转录沉默的GUS标记基因的表达。为了获得高效的GUS扇形化，构建了在热激诱导型启动子控制下携带FLP重组酶的单插入拟南芥株系以及一个FLP可激活的GUS构建体。在不同发育阶段给予各种热激处理后，对两个插入片段均为半合子和纯合子的株系中的GUS扇形化进行了分析。结果表明，在拟南芥的大多数营养、地上和有性结构中，GUS扇形化事件能够被有效诱导。此外，通过选择热激的条件和时间，扇形化事件的频率、扇形大小以及在一定程度上产生扇形的组织都能够很容易地得到控制。
