Cloning of expansin genes in ramie (Boehmeria nivea L.) based on universal fast walking

Jie Chen; Lunjin Dai; Bo Wang; Lijun Liu; Dingxiang Peng

doi:10.1016/j.gene.2014.11.029

Cloning of expansin genes in ramie (Boehmeria nivea L.) based on universal fast walking

Gene. 2015 Sep 10;569(1):27-33. doi: 10.1016/j.gene.2014.11.029. Epub 2014 Dec 4.

Authors

Jie Chen¹, Lunjin Dai², Bo Wang³, Lijun Liu⁴, Dingxiang Peng⁵

Affiliations

¹ MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, #1 Shizishan Street, Hongshan District, Wuhan 430070, Hubei Province, China. Electronic address: lqlcj@126.com.
² MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, #1 Shizishan Street, Hongshan District, Wuhan 430070, Hubei Province, China. Electronic address: dailunjin@gmail.com.
³ MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, #1 Shizishan Street, Hongshan District, Wuhan 430070, Hubei Province, China. Electronic address: wangbo@mail.hzau.edu.cn.
⁴ MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, #1 Shizishan Street, Hongshan District, Wuhan 430070, Hubei Province, China. Electronic address: liulijun@mail.hzau.edu.cn.
⁵ MOA Key Laboratory of Crop Ecophysiology and Farming System in the Middle Reaches of the Yangtze River, College of Plant Science and Technology, Huazhong Agricultural University, #1 Shizishan Street, Hongshan District, Wuhan 430070, Hubei Province, China. Electronic address: pdxiang@mail.hzau.edu.cn.

PMID: 25481635
DOI: 10.1016/j.gene.2014.11.029

Abstract

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 α-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.

Keywords: Degenerate primer; Expansin; Gene cloning; Ramie; Touchdown PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Boehmeria / genetics*
Cloning, Molecular / methods*
DNA, Plant
Gene Expression Regulation, Plant
Molecular Sequence Annotation*
Plant Proteins / genetics*
Plant Proteins / isolation & purification

Substances

DNA, Plant
Plant Proteins
expansin protein, plant