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Tissue-specific and developmental regulation of cotton gene FbL2A. Demonstration of promoter activity in transgenic plants.

机译:棉花基因FbL2A的组织特异性和发育调控。证明了转基因植物中启动子的活性。

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摘要

A gene (FbL2A) that is preferentially expressed in cotton (Gossypium barbadense L. cv Sea Island) fiber was isolated and characterized. Genomic and cDNA analyses suggest multiple FbL2A genes in cotton. The gene is developmentally regulated and is activated during late primary and early secondary wall synthesis stages. FbL2A encodes a polypeptide of 43.4 kD and a predicted isoelectric point of 5.97. The nucleotide-derived protein is highly hydrophilic except for a hydrophobic N terminus and has a compositional bias for glutamic acid (26.3 mol%) and lysine (18.9 mol%). Sixty-two percent of the putative protein is composed of repeat motifs. A 55-amino-acid peptide region is repeated four times in a concatenate fashion within the protein. The function of the protein in the fiber cells is not known. A 2.3-kb DNA fragment 5' from the FbL2A gene is shown to direct expression of heterologous proteins in transgenic cotton in a fiber-specific and developmentally regulated fashion. The FbL2A promoter was used to express in transgenic cotton genes encoding acetoacetyl-coenzyme A reductase and polyhydroxyalkanoic acid synthase, which are involved in the synthesis of the thermoplastic polymer polyhydroxybutyric acid. Transgenic plants containing both enzymes produced polyhydroxybutyric acid in fiber. Thus, the FbL2A promoter is useful in genetic engineering schemes to modify cotton fiber.
机译:分离并鉴定了优先在棉花(巴贝斯棉(L. cv)海岛棉)纤维中表达的基因(FbL2A)。基因组和cDNA分析表明棉花中存在多个FbL2A基因。该基因受到发育调控,并在初级和次级晚期壁合成阶段被激活。 FbL2A编码的多肽为43.4 kD,预测的等电点为5.97。核苷酸衍生的蛋白质除疏水性的N末端外具有高度亲水性,并且对谷氨酸(26.3mol%)和赖氨酸(18.9mol%)具有组成偏倚。假定蛋白质的62%由重复基序组成。 55个氨基酸的肽区域以串联方式在蛋白质中重复四次。蛋白质在纤维细胞中的功能尚不清楚。 FbL2A基因的2.3 kb DNA片段5'以纤维特异性和发育调控的方式指导异源蛋白质在转基因棉花中的表达。 FbL2A启动子用于在转基因棉花中表达编码乙酰乙酰辅酶A还原酶和聚羟基链烷酸合酶的基因,这些基因参与热塑性聚合物聚羟基丁酸的合成。包含这两种酶的转基因植物在纤维中产生了聚羟基丁酸。因此,FbL2A启动子可用于基因工程计划以修改棉纤维。

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