title: Cloning and expression characteristics of the notch-associated gene BmE(spl)mγ from silkworm, Bombyx mori.
abstract: The E(spl)mγ gene in Drosophila is a regulatory target gene downstream of the Notch pathway. BmE(spl)mγ (Bombyx mori, E(spl)mγ) is an ortholog of the Drosophila E(spl)mγ gene, and the gene encodes a protein with 248 amino acid residues. This gene was cloned and overexpressed in Escherichia coli BL21(DE3). The recombinant protein was purified and subsequently used to generate a rabbit polyclonal antibody. Western blotting analyses showed that BmE(spl)mγ expression is high in pupa and egg, and low in larva and moth. In the fifth instar larva, the protein levels are high in head, epidermis, sexual gland, trachea, and the fatbody and low in the Malpighian tubule, hemolymph, gut, and silk gland. The further immunohistochemical analyses also showed higher BmE(spl)mγ expression in the head of fifth instar larva and pupa. Of the four moth parts studied, the thorax had the highest expression level. Thus, BmE(spl)mγ might be associated with neurogenesis in silkworm. Furthermore, DAPT (a γ-secretase inhibitor and an indirect inhibitor of Notch) blocking experiments showed that higher concentrations of the blocking agent and a longer processing time reduce the transcription levels of the BmE(spl)mγ gene, demonstrating that the silkworm BmE(spl)mγ gene is associated with the Notch signal pathway. These findings suggest that the function of BmE(spl)mγ may be similar to that of its Drosophila homolog.
authors.affiliation: College of Life Sciences, Zhejiang Sci-Tech University, Hanghzou, 310018, China.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.forename: Chan
authors.initials: C
authors.lastname: Wang
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.forename: Yue
authors.initials: Y
authors.lastname: Liu
authors.forename: Qing
authors.initials: Q
authors.lastname: Sheng
authors.forename: Zhengbing
authors.initials: Z
authors.lastname: Lv
authors.forename: Wei
authors.initials: W
authors.lastname: Yu
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.forename: Yaozhou
authors.initials: Y
authors.lastname: Zhang
authors.forename: Zuoming
authors.initials: Z
authors.lastname: Nie
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D012831:Bombyx / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D021381:Protein Transport; D011679:Pupa / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D011817:Rabbits; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction
chemical_list: D019476:Insect Proteins; D051880:Receptors, Notch; D012097:Repressor Proteins