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Sequence
analysis and over-expression of ribosomal protein S28 gene (RPS28)
from the Giant Panda
Wan-ru Hou1*,
Yi-ling Hou1, Yan-zhe Hao1, Yu-jie Du2,
Tian-Zhang1 and Zheng-song Peng1
1College
of Life Science, China West Normal University, 44# Yuying
Road, 637002, Nanchong, China.
2Zhan
Jiang educational College, Zhanjiang, China.
*Corresponding author. E-mail:
hwr168@yahoo.com.cn.
Phone/Fax: +86-0817-2568653.
Accepted 30 January, 2009 |
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RPS28 is a component of the
40S small ribosomal subunit
encoded by RPS28 gene, which is
specific to eukaryotes.
The cDNA and the genomic sequence of
RPS28 were
cloned successfully
from the Giant Panda using RT-PCR technology and Touchdown-PCR,
respectively. Both sequences were analyzed preliminarily and
the cDNA of the RPS28 gene was also overexpressed in
Escherichia coli ER2566 and immunoblotted. The cDNA
of the RPS28 cloned from Giant Panda is 216 bp in
size, containing an open reading frame of 210 bp encoding 69
amino acids. The length of the genomic sequence is 790 bp,
which was found to possess three exons and two introns.
Alignment analysis indicated that the nucleotide sequence of
the coding sequence shows a high homology to those of
Homo sapiens, Bos Taurus, Mus musculus,
Rattus norvegicus and
Sus scrofa (92.4, 92.4, 87.1, 86.7 and 89.5%, respectively) as
determined by Blast analysis.. The amino acid sequence
encoded by RPS28 gene of the Giant Panda shared a
high homology (100%) with those of Homo sapiens,
B. Taurus, M. musculus, R. norvegicus and
S. scrofa. Primary structure analysis revealed that the molecular weight of the
putative RPS28 protein is 7.841 kD with a theoretical pI
10.70. Topology prediction showed there is one ribosomal
protein S28e signature site, three protein kinase C
phosphorylation site and two casein kinase II
phosphorylation site in the RPS28 protein of the Giant
Panda. The RPS28 gene can be really expressed in
Escherichia coli and the RPS28 protein fusioned with the
N-terminally GST -tagged protein gave rise to the
accumulation of an expected 34 KDa polypeptide. The
expression product obtained could be used for purification
and study of its function further.
Key words: RPS28
gene,
ribosomal protein S28 (RPS28),
giant panda, (Ailuropoda melanoleuca), genomic
cloning, overexpression. |
