棉花种子特异性启动子的克隆及序列分析

启动子是基因表达的重要顺式调控元件,在基因工程中,种子特异性启动子可以调控外源基因在种子中特异表达,提高表达效率,增强转基因的效果。本试验根据棉花LEA蛋白D34基因序列设计引物,以海岛棉基因组DNA为模板,通过touch down PCR技术,获得D34基因的种子特异性启动子片段。测序结果表明该片段长1 384 bp,与已发表的D34基因序列相似度为94.87%。该片段除了含有启动子的基本元件TATA框、CAAT框外,还含有种子特异性启动子元件E-box、G-box、B-box、AACA基序等。此外,对其顺式元件做了生物学功能分析。

Cloning and Sequence Analysis of Seed-Specific Promoter in Cotton

Promoter is a crucial cis-regulatory element of gene expression .In genetic engineering , seed-specific promoter plays an important role in regulating the specific expression of exogenous genes in seeds , im-proving the expression efficiency and enhancing the transgenic effect .In this paper , a seed-specific promoter sequence of gene D34 was cloned from Gossypium barbadense by touch down PCR amplification with the prim-ers designed based on the D34 gene of cotton LEA protein .Sequence analysis showed that the cloned fragment was 1 384 bp in length and similar to published sequence with the similarity as 94 .87%.The cloned segment not only contained promoter basic components such as TATA box , CAAT box, but also contained seed -spe-cific promoter components such as E -box , G-box and AACA motif .The biological function of these cis -el-ements was also analyzed .