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Expression analysis and
characteristics of Bm-LOC778477 gene, a hypothetical protein
from silkworm pupae, Bombyx mori
Wei Yu, Aiwu Tan and Yaozhou Zhang*
Institute
of Biochemistry, College of life science, Zhejiang Sci-Tec
University, Hangzhou 310018, China.
*Corresponding author.
E-mail:
yaozhou@zstu.edu.cn. Tel: +86-571-86843199. Fax:
+86-571-86843198.
Accepted 2
June, 2009 |
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This is the first identification of a gene encoding
Bm-LOC778477 protein (a “hypothetical protein”) from
silkworm pupal cDNA library. The full-length Bm-LOC778477
cDNA contained a 546-bp ORF encoding 187 amino acids. Using
bioinformatics, the Bm-LOC778477 gene was predicted
on the W chromosome and 4 exons and 3 introns were
determined when compared to the silkworm genome. The
consensus splice sites of exon/intron junctions were
consistent with Chambon′s rule. In addition, TATA box and
initiator (Inr) were also predicated in the promoter region
of the gene. To elucidate whether Bm-LOC778477 gene encoded
a functional protein in silkworm, pGEX-4T-1-Bm-LOC778477
expression plasmid was constructed to express GST-LOC778477
fusion protein in Escherichia coli Rosetta.
rBm-LOC778477 was used to generate anti- Bm-LOC778477
polyclonal antibody, which were used to determine the
subcellular localization of Bm-LOC778477. Immunostaining
indicated that Bm-LOC778477 protein could be found in both
the cytoplasm and nucleus. Western blot analysis indicated
that Bm-LOC778477 was specific expression during the pupa
stage and there was seldom expression during the larvae
stage. Therefore, we propose that Bm-LOC778477 may play an
important role during pupa stage development. These results
may lay an important foundation of further function studies
of Bm-LOC778477 protein.
Key
words:
Bm-LOC778477, hypothetical protein, subcellular
localization. |
