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首页> 外文期刊>Indian Phytopathology >Isolation and characterization of partial secA gene from coconut root wilt and arecanut yellow leaf disease phytoplasma and assessment of its possible use for sero diagnosis
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Isolation and characterization of partial secA gene from coconut root wilt and arecanut yellow leaf disease phytoplasma and assessment of its possible use for sero diagnosis

机译：椰子根枯萎病和槟榔黄叶病植物原质中secA基因的部分分离，鉴定及其在血清学诊断中的应用价值

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摘要

A 425 bp fragment of phytoplasma secA gene encoding SecA, a component of the Sec protein translocation system, was isolated and characterized from coconut root wilt disease and arecanut yellow leaf disease phytoplasma, both belonging to the 16SrXI-B sub group phytoplasma.The gene showed 99% nucleotide identity with sugarcane grassy shoot phytoplasma secA gene. In the phylogenetic analysis of the translated SecA protein sequence, all the 16SrXI group phytoplasmas clustered together. The three dimensional structure of the protein was predicted using Moddler 9.13. The validation of the predicted structure indicated that 83.6% of the amino acids lie in the favored region indicating that the predicted structure was ideal. The possible antigenic domains in the protein were also predicted using the tool SVM TriP. The antigenicity prediction of the SecA protein fragment showed the presence of 4 possible antigenic domains.The secA gene was expressed in pET vector and the protein was used for the developmentof polyclonal antiserum in rabbit. Because the titre of the phytoplasma was very less, the color development in ELISA was less intense. A technique for isolation of higher number of phytoplasma cells has to be standardized.
机译：从属于16SrXI-B亚类植物质体的椰子根枯萎病和槟榔黄叶病植物质体中分离并鉴定了编码SecA的植物质体secA基因的425 bp片段，并对其进行了鉴定。与甘蔗草芽植物质体secA基因具有99％的核苷酸同一性。在翻译的SecA蛋白序列的系统发育分析中，所有16SrXI组植原体都聚集在一起。使用Moddler 9.13预测了蛋白质的三维结构。对预测结构的验证表明，有83.6％的氨基酸位于有利区域中，表明预测结构是理想的。还使用工具SVM TriP预测了蛋白质中可能的抗原结构域。 SecA蛋白片段的抗原性预测表明存在4个可能的抗原结构域。secA基因在pET载体中表达，该蛋白用于兔多克隆抗血清的开发。由于植物原浆的滴度非常小，因此ELISA中的显色性较弱。分离更多数量的植物浆细胞的技术必须标准化。

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