Cloning of expansin genes in ramie (Boehmeria nivea L.) based on universal fast walking
Cloning of expansin genes in ramie (Boehmeria nivea L.) based on universal fast walking
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DOI：
10.1016/j.gene.2014.11.029
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发表时间：
2015-09-10
期刊：
影响因子：
3.5
通讯作者：
Peng, Dingxiang
中科院分区：
文献类型：
--
作者：
Chen, Jie;Dai, Lunjin;Peng, Dingxiang
基因克隆是研究特定基因的表达谱和功能的第一步;已经开发了相当多的克隆方法。苎麻（Boehmeria nivea L.）是重要的韧皮纤维作物之一，其Expansin基因被认为参与了细胞壁的修饰，但在苎麻中尚未克隆到。对expansin基因家族的研究将揭示其与苎麻纤维发育及其他生长事件的可能关系。结果，获得了5个expansin基因的全长序列，并对其序列进行了分析。系统发育分析表明，克隆的基因属于α-亚家族，且在苎麻纤维发育过程中表达差异。本研究利用简并引物、降落PCR和通用快速步移法从苎麻基因组DNA中克隆全长新基因。通过克隆5个expansin基因的全长，我们认为基于聚合酶链反应的基因克隆策略可以应用于苎麻和其他作物的一般基因研究。(C)2014由Elsevier B. V.出版
Gene cloning is the first step to study the expression profiles and functions of a particular gene; considerable cloning methods have been developed. Expansin, thought to involve in the cell-wall modification events, was not cloned in ramie (Boehmeria nivea L), which is one of the most important bast fiber crops with little conducted molecular research, especially on its fiber development. Studying the expansin gene family will uncover its possible relationship with ramie fiber development and other growth events. As a result, five expansin genes were cloned with full-length and their sequence information was investigated. Additionally, the phylogenetic analysis was conducted, which suggested that the cloned genes belong to the a-subfamily, and these genes expressed differently during ramie fiber developmental process. In this study, we aimed to apply a strategy for cloning novel full-length genes from genomic DNA of ramie, based on using degenerate primers, touchdown polymerase chain reaction and universal fast walking protocols. By cloning five full-length expansin genes, we believe the polymerase chain reaction-based gene cloning strategy could be applied to general gene studies in ramie and other crops. (C) 2014 Published by Elsevier B.V.