Pauls P K, Kunert K, Huttner E, Grandbastien M A
Laboratoire de Biologie Cellulaire, INRA-Versailles, France.
Plant Mol Biol. 1994 Oct;26(1):393-402. doi: 10.1007/BF00039548.
The expression of the tobacco (Nicotiana tabacum) retrotransposon Tnt1 has previously been shown to be strongly regulated and driven from the 5' long terminal repeat (LTR). We report here that the Tnt1 LTR can promote activity of the beta-glucuronidase (GUS) reporter gene in two heterologous species of the Brassicaceae family, namely rapeseed (Brassica napus) and Arabidopsis thaliana. The translational LTR-GUS fusion was active in transient expression studies performed with tobacco and rapeseed protoplasts, indicating that the LTR sequences are recognized in heterologous species. Our results also showed that Tnt1 LTR-promoted GUS expression in transgenic Arabidopsis is strongly regulated, and that, in contrast to tobacco, hormonal activation plays a significant role in the expression of the Tnt1 LTR in Arabidopsis. LTR sequences were shown to be more effective than the CaMV 35S enhancer region in transient expression studies performed with tobacco or rapeseed protoplasts, and substitution of the LTR sequences upstream from the major transcriptional start with the CaMV 35S enhancer region gave high levels of expression in transgenic tobacco and Arabidopsis leaves, suggesting that a Tnt1 element with similar substitutions in its 5' LTR might be suited for gene-tagging experiments in heterologous species.
烟草(Nicotiana tabacum)反转录转座子Tnt1的表达先前已被证明受到严格调控,并由5'长末端重复序列(LTR)驱动。我们在此报告,Tnt1 LTR可促进十字花科两个异源物种即油菜(Brassica napus)和拟南芥(Arabidopsis thaliana)中β-葡萄糖醛酸酶(GUS)报告基因的活性。翻译LTR-GUS融合体在烟草和油菜原生质体进行的瞬时表达研究中具有活性,表明LTR序列在异源物种中可被识别。我们的结果还表明,Tnt1 LTR促进的GUS在转基因拟南芥中的表达受到严格调控,并且与烟草不同,激素激活在拟南芥中Tnt1 LTR的表达中起重要作用。在烟草或油菜原生质体进行的瞬时表达研究中,LTR序列比CaMV 35S增强子区域更有效,并且用CaMV 35S增强子区域替换主要转录起始上游的LTR序列在转基因烟草和拟南芥叶片中产生了高水平的表达,这表明在其5' LTR中具有类似替换的Tnt1元件可能适用于异源物种中的基因标签实验。