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

J. Li; S. L. Wang; M. Cao; D. W. Lv; S. Subburaj; X. H. Li; F. J. Zeller; S. L.K. Hsam; Y. M. Yan

doi:10.1007/s13353-013-0139-z

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

J. Li, S. L. Wang, M. Cao, D. W. Lv, S. Subburaj, X. H. Li, F. J. Zeller, S. L.K. Hsam, Y. M. Yan

Chair of Plant Breeding

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Fifteen novel α-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 α-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these α-gliadins to be assigned to specific chromosomes. Five representative α-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-β-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T₇ promoter. The transcriptional expression of α-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) α-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that α-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.

Original language	English
Pages (from-to)	157-167
Number of pages	11
Journal	Journal of Applied Genetics
Volume	54
Issue number	2
DOIs	https://doi.org/10.1007/s13353-013-0139-z
State	Published - May 2013

Keywords

Heterologous expression
Phylogenetic evolution
Quantitative real-time PCR
Toxic epitopes
α-Gliadins

Access to Document

10.1007/s13353-013-0139-z

Cite this

@article{c43137905fb947f5a2ced65ae5821697,

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

abstract = "Fifteen novel α-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 α-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these α-gliadins to be assigned to specific chromosomes. Five representative α-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-β-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T7 promoter. The transcriptional expression of α-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) α-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that α-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.",

keywords = "Heterologous expression, Phylogenetic evolution, Quantitative real-time PCR, Toxic epitopes, α-Gliadins",

author = "J. Li and Wang, {S. L.} and M. Cao and Lv, {D. W.} and S. Subburaj and Li, {X. H.} and Zeller, {F. J.} and Hsam, {S. L.K.} and Yan, {Y. M.}",

note = "Funding Information: This research was financially supported by grants from the Chinese Ministry of Science and Technology (2009CB118303), the National Natural Science Foundation of China (31271703, 31101145), and the National Key Projects for Transgenic Crops of China (2011ZX08009-003-004, 2011ZX08002-004).",

year = "2013",

month = may,

doi = "10.1007/s13353-013-0139-z",

language = "English",

volume = "54",

pages = "157--167",

journal = "Journal of Applied Genetics",

issn = "1234-1983",

publisher = "Springer Science and Business Media Deutschland GmbH",

number = "2",

}

TY - JOUR

T1 - Cloning, expression, and evolutionary analysis of α-gliadin genes from Triticum and Aegilops genomes

AU - Li, J.

AU - Wang, S. L.

AU - Cao, M.

AU - Lv, D. W.

AU - Subburaj, S.

AU - Li, X. H.

AU - Zeller, F. J.

AU - Hsam, S. L.K.

AU - Yan, Y. M.

N1 - Funding Information: This research was financially supported by grants from the Chinese Ministry of Science and Technology (2009CB118303), the National Natural Science Foundation of China (31271703, 31101145), and the National Key Projects for Transgenic Crops of China (2011ZX08009-003-004, 2011ZX08002-004).

PY - 2013/5

Y1 - 2013/5

N2 - Fifteen novel α-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 α-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these α-gliadins to be assigned to specific chromosomes. Five representative α-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-β-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T7 promoter. The transcriptional expression of α-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) α-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that α-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.

AB - Fifteen novel α-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 α-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these α-gliadins to be assigned to specific chromosomes. Five representative α-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-β-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T7 promoter. The transcriptional expression of α-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) α-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that α-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.

KW - Heterologous expression

KW - Phylogenetic evolution

KW - Quantitative real-time PCR

KW - Toxic epitopes

KW - α-Gliadins

UR - http://www.scopus.com/inward/record.url?scp=84876035648&partnerID=8YFLogxK

U2 - 10.1007/s13353-013-0139-z

DO - 10.1007/s13353-013-0139-z

M3 - Article

C2 - 23456845

AN - SCOPUS:84876035648

SN - 1234-1983

VL - 54

SP - 157

EP - 167

JO - Journal of Applied Genetics

JF - Journal of Applied Genetics

IS - 2

ER -

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

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this