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Plant Gene and Trait 2012, Vol.3, No.1, 1
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Research Report Open Access
Cloning and Sequence Analysis of Actin Gene from
Guzmania
Jianxin Liu , Yaying Ge
,
Danqing Tian
,
Zhi Zhang
,
Huaqiao Ding
,
Fuquan Shen
,
Weiyong Wang
Research & Development Centre of Flower, Zhejiang Academy of Agricultural Sciences, Hangzhou, 311202, P.R China
Corresponding author email:
ljxljx20002000@yahoo.com.cn;
Authors
Plant Gene and Trait, 2012, Vol.3, No.1 doi: 10.5376/pgt.2012.03.0001
Received: 05, Nov., 2010
Accepted: 30, Nov., 2011
Published: 05, Dec., 2011
This article was first published in Molecular Plant Breeding (Regular Print Version) in Chinese, and here was authorized to redistribute in English under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Liu et al., 2011, Cloning and sequence analysis of
Actin
gene from
Guzmania
, Plant Gene and Trait, Vol.3, No.1 1-5 (doi: 10.5376/pgt.2012.03.0001)
Abstract
As a house keeping gene,
Actin
was often used as inner control in quantitative and semiquantitative PCR tests to
determine relative expressive amount of target genes. Based on EST monoclones of
Actin
gene obtained from
Guzmania
full length
cDNA library, its full length cDNA sequence was obtained by primer walking sequencing. The gene, named for
Goactin1
(GenBank
accession No. HQ184438), consisted of 1 625 bp cDNA sequence, 1 131 bp ORF (open reading frame) which encoded a protein with
377 amino acids residues, and a putative protein which was 41.7 kD at estimated molecular weight, 5.31 at isoelectric point and had a
‘actin superfamily’ conservative domain. The secondary structure of the protein was composed of random coil, alpha helix, extend
strand and beta turn by SPOMA analysis. Furthermore, its tertiary structure was also build based on A chain of 2BTF. Through
phylogenetic tree analysis,
Goactin1
was gathered to the same group with Actin protein in
Solanum tuberosum
,
Gossypium hirsutum,
Nicotiana tabacum, Brachypodium sylvaticum, and Arabidopsis thaliana
.
Keywords
Guzmania
(
Guzmania Ruiz&Pav
);
Actin
; Clone
Background
Plant actin was a kind of constitutive expression
protein existed broadly, and had high conservatism
(Meagher et al., 1999). Since Yan and Shi put forward
that actin existed in higher plant for the first time in
1963(Yan and Shi, 1963), at present, actin had been
discovered in pollen, root hair, stem cambium layer,
stem phloem, vagina cell, leaf epidermal cell, tendril
and endocarp tissues of higher plant (Kost et al., 1999),
and also was multi-gene family like as that of animal
and fungi (Zhang and Liu, 2006; Hussey et al., 2006).
Plant actin involved in cell microfilaments formation,
whereas, microfilaments were key components of
cytoskeleton and participated in important physiological
activities such as intracellular cytoplasmic streaming,
keep of configuration, movement, division, differentiated,
transport of substance, signal transduction and polarity
constructing etc. (Ma et al., 2010). As house-keeping
gene,
Actin
can express relative stably to some extent
in tissues and cells and had many advantages such as
constitutive expression, multiplication easily. Hence,
like as other house-keeping genes included
GAPDH,
18SrRNA, TBP and
β
-2-microglobulin, Actin
often
was used as inner reference in quantitative or
semiquantitative PCR reaction to revise expression
amount of target genes (Zhu et al., 2006; Yang et al.,
2007; Liu, 2003).
Guzmania
, belonged to
Bromeliaceae
, was native to
tropics and suibtropics of American, and was superior
quality potted flower plant in flower marketplace. At
present, there was a lack of information in public
about
Guzmania
Actin
gene. Based on full length
cDNA library constructed successfully in
Guzmania
ostara
and plenty of EST sequences information were
obtained by sequencing randomly on a large scale
formerly, full length cDNA sequence of
Actin
gene
was obtained, by Primer Walking sequencing to EST
monoclones belonged to
Actin
gene. Finally, there was
a bioinformatics analysis in-depth to the gene. The
research laid a foundation to construction of quantitative
and semiquantitative PCR technique in ornamental
Bromeliaceae.
1 Results and Analysis
1.1 Cloning of
Goactin1
gene
Through primer walking sequencing to EST monoclone:
|ppfca0_001242.z1.scf|, which belonged to
Actin
gene,