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一种植物组蛋白基因启动子可在转基因植物中指导依赖复制和不依赖复制的基因表达。

A plant histone gene promoter can direct both replication-dependent and -independent gene expression in transgenic plants.

作者信息

Lepetit M, Ehling M, Chaubet N, Gigot C

机构信息

Biology Department, Texas A & M University, College Station.

出版信息

Mol Gen Genet. 1992 Jan;231(2):276-85. doi: 10.1007/BF00279801.

Abstract

Chimeric genes containing the beta-glucuronidase (GUS) gene under the control of different Arabidopsis histone H3 and H4 promoters were found to be highly expressed in transient expression experiments using tobacco protoplasts. The activity of one of these promoters, H4A748, was further analyzed. The kinetics of H4A748-GUS activity are very similar to these of a CaMV 35S-GUS constitutive gene during protoplast culture. No increase in H4A748-GUS activity was found after 24 h of protoplast culture when DNA synthesis starts, nor was the GUS activity affected when an inhibitor of DNA synthesis was included in the culture medium. This failure to detect any replication-dependent activity is most likely to be due to the fact that transient transcription of the introduced construct is restricted to the first 24 h following transfection. Stable integration of the H4A748-GUS gene into tobacco plants showed that the histone promoter could confer increased expression in meristematic tissues but it is also expressed to significant levels in non-proliferating tissues. Protoplasts prepared from these transgenic tobacco plants were cultivated under different conditions that affect DNA synthesis. Analysis of H4A748-GUS activity revealed (i) the existence of a basal replication-independent activity and (ii) a replication-dependent activity induced in parallel with DNA synthesis. These results show that the histone H4 promoter is able to direct both replication-dependent and -independent gene expression.

摘要

在使用烟草原生质体的瞬时表达实验中,发现含有在不同拟南芥组蛋白H3和H4启动子控制下的β-葡萄糖醛酸酶(GUS)基因的嵌合基因高度表达。对其中一个启动子H4A748的活性进行了进一步分析。在原生质体培养过程中,H4A748-GUS活性的动力学与CaMV 35S-GUS组成型基因的动力学非常相似。在原生质体培养24小时DNA合成开始后,未发现H4A748-GUS活性增加,当培养基中加入DNA合成抑制剂时,GUS活性也未受影响。未能检测到任何依赖复制的活性很可能是由于导入构建体的瞬时转录仅限于转染后的前24小时。将H4A748-GUS基因稳定整合到烟草植株中表明,组蛋白启动子可在分生组织中赋予增强的表达,但在非增殖组织中也有显著表达。从这些转基因烟草植株制备的原生质体在影响DNA合成的不同条件下培养。对H4A748-GUS活性的分析揭示了(i)存在一种基础的不依赖复制的活性和(ii)一种与DNA合成平行诱导的依赖复制的活性。这些结果表明,组蛋白H4启动子能够指导依赖复制和不依赖复制的基因表达。

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