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Construction and analysis of EST libraries of the trans-polyisoprene producing plant, Eucommia ulmoides Oliver

机译:反式聚异戊二烯生产植物杜仲(Eucommia ulmoides Oliver)EST库的构建和分析

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Eucommia ulmoides Oliver is one of a few woody plants capable of producing abundant quantities of trans-polyisoprene rubber in their leaves, barks, and seed coats. One cDNA library each was constructed from its outer stem tissue and inner stem tissue. They comprised a total of 27,752 expressed sequence tags (ESTs) representing 10,520 unigenes made up of 4,302 contigs and 6,218 singletons. Homologues of genes coding for rubber particle membrane proteins that participate in the synthesis of high-molecular poly-isoprene in latex were isolated, as well as those encoding known major latex proteins (MLPs). MLPs extensively shared ESTs, indicating their abundant expression during trans-polyisoprene rubber biosynthesis. The six mevalonate pathway genes which are implicated in the synthesis of isopentenyl diphosphate (IPP), a starting material of poly-isoprene biosynthesis, were isolated, and their role in IPP biosynthesis was confirmed by functional complementation of suitable yeast mutants. Genes encoding five full-length trans-isoprenyl diphosphate synthases were also isolated, and two among those synthesized farnesyl diphosphate from IPP and dimethylallyl diphosphate, an assumed intermediate of rubber biosynthesis. This study should provide a valuable resource for further studies of rubber synthesis in E. ulmoides.
机译:杜仲(Eucommia ulmoides)Oliver是少数能在其叶片,树皮和种皮中产生大量反式聚异戊二烯橡胶的木本植物之一。从其外部茎组织和内部茎组织分别构建一个cDNA文库。它们总共包含27,752个表达序列标签(EST),代表10,520个单基因,由4,302个重叠群和6,218个单例组成。分离出编码参与乳胶中高分子聚异戊二烯合成的橡胶颗粒膜蛋白的基因的同源物,以及编码已知主要乳胶蛋白(MLP)的基因。 MLP广泛共享EST,表明它们在反式聚异戊二烯橡胶生物合成过程中大量表达。分离了与聚异戊二烯生物合成的起始原料异戊二烯二磷酸(IPP)的合成有关的六个甲羟戊酸途径基因,并通过合适的酵母突变体的功能互补证实了它们在IPP生物合成中的作用。还分离了编码五个全长反式异戊二烯基二磷酸合酶的基因,其中两个是从IPP和假定的橡胶生物合成中间体二磷酸二烯丙基二磷酸酯合成的法呢基二磷酸。该研究应为进一步研究大肠埃希氏菌中的橡胶合成提供有价值的资源。

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