Open AccessExpression and subcellular localization of a novel gene <i>Bm-X</i> of the silkworm, <i>Bombyx mori</i>
Author(s) -
Zhengbing Lv,
Yunlong Liu,
Ying Chen,
Bo Li,
Huan Liu,
Jian Chen,
Zhou Nie,
Qing Sheng,
Wengong Yu,
Yaozhou Zhang
Publication year - 2012
Publication title -
american journal of molecular biology
Language(s) - English
Resource type - Journals
eISSN - 2161-6663
pISSN - 2161-6620
DOI - 10.4236/ajmb.2012.23023
Subject(s) - complementary dna , microbiology and biotechnology , biology , bombyx mori , subcellular localization , gene , fusion protein , cytoplasm , western blot , gene expression , cdna library , bombyx , polyclonal antibodies , biochemistry , recombinant dna , antibody , genetics
According to the large-scale sequencing of cDNA library from silkworm pupae, the cDNA of a novel gene with blank research background was identified and temporarily named Bm-X. The length of this cDNA is 778 bp. We obtained its ORF for further study by bioinformatics analysis. It is 444 bp and encodes 147 amino acid residues, with a predicted molecular weight (MW) of 16.4 kD and an isoelectric point (pI) of 3.69. In this study, we successfully constructed a recombinant plasmid pET-28a(+)-Bm-X and expressed it in Escherichia coli. We used the fusion protein rBm-X which purified by Niaffinity chromatography to produce polyclonal antibodies against Bm-X for Western blot analysis. The analysis revealed that Bm-X was expressed in the larval midgut, the epidermis and the silk gland. In addition, the subcellular localization analysis of silkworm ovary epithelial cells (BmN cells) showed that Bm-X protein was located both in cytoplasm and nucleus, and the signal was stronger in cytoplasm than in nucleus. Our findings indicate that Bm-X gene is a novel species-specificity gene and its expression product can be detected in tissues of the fifth silkworm instar larvae and BmN cells