Abstract
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.
Highlights
Silkworm (Bombyx mori) is a holometabolic insect which has complete biology background and is a model organism of lepidoptera insects
Using specific polymerase chain reaction (PCR) primers on our in-house plasmid TG1-pHelix-Bm-X, we amplified the cDNA of the B. mori-X gene (Bm-X)
Characteristics of Deduced Bm-X Protein by Bioinformatics Analysis BLASTX analysis of the translation product deduced from the open reading frame (ORF) showed that Bm-X has no significant homology with other proteins in the GenBank database
Summary
Silkworm (Bombyx mori) is a holometabolic insect which has complete biology background and is a model organism of lepidoptera insects. The silkworm genome consists of 28 pairs of chro-. After complete sequence determination and analysis of silkworm genome, more than 160,000 complete genes and 7000 gene fragments were obtained. Silkworm has more than 20,000 genes and over 25% of them are novel genes [2,3]. We have constructed a silkworm pupae cDNA library from which isolated 2400 cDNA sequences were isolated. Our library contains many novel genes of unknown function [4]. In our efforts to clarify these unknown genes, we have previously reported the selective expression of the silkworm troponin C gene during silkworm development [5]. Wang demonstrated that the expression of BmCRABP (silkworm Cellular retinoic acid binding protein) gene had effect on the physiological function of atRA [6]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
