Cloning, expression, and evolutionary analysis of α-gliadin genes from Triticum and Aegilops genomes

被引：11
|
作者：
Li, J. [1 ]
Wang, S. -L. [1 ]
Cao, M. [1 ]
Lv, D. -W. [1 ]
Subburaj, S. [1 ]
Li, X. -H. [1 ]
Zeller, F. J. [2 ]
Hsam, S. L. K. [2 ]
Yan, Y. -M. [1 ]
机构：
[1] Capital Normal Univ, Coll Life Sci, Beijing 100048, Peoples R China
[2] Tech Univ Munich, D-85354 Freising Weihenstephan, Germany
来源：
JOURNAL OF APPLIED GENETICS | 2013年 / 54卷 / 02期
基金：
中国国家自然科学基金;
关键词：
alpha-Gliadins; Toxic epitopes; Heterologous expression; Quantitative real-time PCR; Phylogenetic evolution; HORDEUM-CHILENSE ROEM; GLUTENIN SUBUNIT GENE; MOLECULAR CHARACTERIZATION; CELIAC-DISEASE; BREAD WHEAT; HETEROLOGOUS EXPRESSION; ALLELIC VARIATION; STORAGE PROTEINS; DOUGH STRENGTH; BETA-TYPE;
D O I：
10.1007/s13353-013-0139-z
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Fifteen novel alpha-gliadin genes were cloned and sequenced from Triticum and related Aegilops genomes by allele-specific polymerase chain reaction (AS-PCR). Sequence comparison displayed high diversities in the alpha-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these alpha-gliadins to be assigned to specific chromosomes. Five representative alpha-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-beta-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T-7 promoter. The transcriptional expression of alpha-gliadin genes during grain development detected by quantitative real-time polymerase chain reaction (qRT-PCR) showed a similar up-down regulation pattern in different genotypes. A neighbor-joining tree constructed with both full-open reading frame (ORF) alpha-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that alpha-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.
引用
收藏
页码：157 / 167
页数：11
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