Basic HTML Version
Plant Gene and Trait 2012, Vol.3, No.5, 22
-
27
http://pgt.sophiapublisher.com
Research Report Open Access
Molecular Cloning of Promoter of
AP3
Gene from
Arabidopsis thaliana
Ecotype
(Col) and Construction of the Plant Expression Vector
Bingyou Fan
1
, Shuiping Gao
2
, Xiaogai Hou
1
, Huawei Xu
1
, Guoan Shi
1
, Xiangsheng Kong
1
1.
College of Agriculture, Henan University of Science and Technology, Luoyang, Henan, 471003, P.R. China
2. College of Forestry, Henan University of Science and Technology, Luoyang, Henan, 471003, P.R. China
Corresponding author email:
fanbingyou2005@163.com;
Authors
Plant Gene and Trait, 2012, Vol.3, No.5 doi: 10.5376/pgt.2012.03.0005
Received: 30 Jan., 2012
Accepted: 24 Feb., 2012
Published: 01 Mar., 2012
This article was first published in Genomics and Applied Biology in Chinese, and here was authorized to translate and publish the paper in English under the terms of 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:
Fan et al., 2012, Molecular Cloning of Promoter of
AP3
Gene from
Arabidopsis thaliana
Ecotype (Col) and Construction of the Plant Expression Vector, Plant Gene and Trait,
Vol.3, No.5 22-27 (doi: 10.5376/pgt.2012.03.0005)
Abstract
Belonging to MADS-Box gene family,
APETALA3
(
AP3
) gene of
Arabidopsis
floral organ class B is expressed in petals
and stamens specifically.
AP3
gene which encodes a transcription factor controls the development of petals and stamens of
dicotyledons by cooperation with class A and class C genes, respectively. The results showed that the promoter of
AP3
was the
flower-specific expression promoter. Therefore, the cloning and functional identification of
AP3
gene promoter from
Arabidopsis
thaliana
will play significant effects on the oriented improvement of the commercial traits related to the flowers of ornamental plants.
Here, a pair of specific PCR primers was designed according to the promoter sequence of
AP3
gene of
Arabidopsis thaliana
ecotype
(Ler) reported in GenBank (U30729). A length of 1 767 bp of
AP3
gene promoter was obtained from
Arabidopsis thaliana
ecotype
(Col) by using PCR technique with high-fidelity DNA polymerase KOD-plus, named as
pAtAP3
, the GenBank accession No. is
FJ619533. Online Bl2seq analysis indicated that the sequence similarity between
pAtAP3
and U30729 was 98%. It also showed that
the sequence similarity between
pAtAP3
and the BAC clone T12E18 (AL132971) of
Arabidopsis thaliana
ecotype (Col) from 9 264
to 11 030 was up to 100%, and its downstream sequence encoded
AP3
protein (CAB81799), which approved that the sequence we
cloned was the promoter of
AP3
gene of
Arabidopsis thaliana
ecotype (Col). Online PLACE analysis displayed that
pAtAP3
contained the basic
cis
-elements of promoter such as TATA-box and CAAT-box. It also comprised several
cis
-elements related to
flower-specific expression, such as CArG1, CArG2, CArG3 and anther-box. Moreover, plant expression vector of
pAtAP3
::
GUS
was
successfully constructed in this research, which would lay the foundation for functional identification of the
AP3
promoter of
Arabidopsis thaliana
ecotype (Col).
Keywords
Arabidopsis thaliana
; Ecotype Col; Promoter of
APETALA3
; Plant expression vector
Background
Based on the study of floral homeotic mutants of
Arabidopsis thaliana
and
Antirrhinum majus
L., Coen
and Meyerowitz (1991) proposed the ABC model of
flower development, subsequently, it was extended to
the ABCD model later by Angenent and Colombo (1996).
In the genetic network of flower development, flower
organ identity genes determine floral organ primordia
developed into sepals, petals, stamens or carpels. The
class B flower organ identity gene,
APETALA3
(AP3
),
encoding transcription factors, belongs to MADS-Box
gene family. Specifically expressed in petal and
stamen,
AP3
gene synergistic controlled the petal and
stamen development of dicotyledons with class A and
class C identity genes (Krizek and Meyerowits, 1996).
Promoter expression analysis displayed that
AP3
promoter was the flower-specific expression promoter
(Verdonk et al., 2008).
When using biotechnology for genetic improvement
on important commercial traits in plant, if the hetero-
logous gene expression was driven by constitutive
promoter such as CaMV 35S, it may inhibit the
growth and development of the transgenic plants.
Under the control of one flower-specific expression
promoter, the foreign gene will be specifically expressed
in floral organs, which will increase the regional
expression, and will not have an adverse development
22