Cloning and promoter analysis of the cotton lipid transfer protein gene Ltp3(1).

Biochimica et biophysica acta Pub Date : 2000-08-24
H C Liu, R G Creech, J N Jenkins, D P Ma
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Abstract

A cotton Ltp3 gene and its 5' and 3' flanking regions have been cloned with a PCR-based genomic DNA walking method. The amplified 2.6 kb DNA fragment contains sequences corresponding to GH3 cDNA which has been shown to encode a lipid transfer protein (LTP3). The gene has an intron of 80 bp which is located in the region corresponding to the C-terminus of LTP3. The Ltp3 promoter was systematically analyzed in transgenic tobacco plants by employing the Escherichia coli beta-glucuronidase gene (GUS) as a reporter. The results of histochemical and fluorogenic GUS assays indicate that the 5' flanking region of the Ltp3 gene contains cis-elements conferring the trichome specific activity of Ltp3 promoter.

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棉花脂质转移蛋白基因 Ltp3(1) 的克隆和启动子分析。
利用基于 PCR 的基因组 DNA 走位法克隆了棉花 Ltp3 基因及其 5' 和 3' 侧翼区域。扩增出的 2.6 kb DNA 片段含有与 GH3 cDNA 相对应的序列,而 GH3 cDNA 被证明编码一种脂质转移蛋白(LTP3)。该基因有一个 80 bp 的内含子,位于与 LTP3 C 端相对应的区域。通过使用大肠杆菌 beta-葡糖醛酸酶基因(GUS)作为报告基因,在转基因烟草植株中对 Ltp3 启动子进行了系统分析。组织化学和荧光 GUS 检测的结果表明,Ltp3 基因的 5' 侧翼区域含有赋予 Ltp3 启动子毛状体特异性活性的顺式元件。
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