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Transgenic characterization of two silkworm tissue‐specific promoters in the haemocyte plasmatocyte cells

Author:
Zhang, K., Li, C., Weng, X., Su, J., Shen, L., Pan, G., Long, D., Zhao, A., Cui, H.
Source:
Insect molecular biology 2018 v.27 no.2 pp. 133-142
ISSN:
0962-1075
Subject:
Bombyx mori, CD18 antigen, Western blotting, fluorescence, genes, genetic engineering, green fluorescent protein, hematopoiesis, hemocytes, innate immunity, metabolism, metamorphosis, models, quantitative polymerase chain reaction, silkworms, transgenic insects
Abstract:
Haemocytes play crucial roles in insect metabolism, metamorphosis, and innate immunity. As a model of lepidopteran insects, the silkworm is a useful model to study the functions of both haematopoiesis and haemocytes. Tissue‐specific promoters are excellent tools for genetic manipulation and are widely used in fundamental biological research. Herein, two haemocyte‐specific genes, Integrin β2 and Integrin β3, were confirmed. Promoter activities of Integrin β2 and Integrin β3 were evaluated by genetic manipulation. Quantitative real‐time PCR and western blotting suggested that both promoters can drive enhanced green fluorescent protein (EGFP) specifically expressed in haemocytes. Further evidence clearly demonstrated that the transgenic silkworm exhibited a high level of EGFP signal in plasmatocytes, but not in other detected haemocyte types. Moreover, EGFP fluorescence signals were observed in the haematopoietic organ of both transgenic strains. Thus, two promoters that enable plasmatocytes to express genes of interest were confirmed in our study. It is expected that the results of this study will facilitate advances in our understanding of insect haematopoiesis and immunity in the silkworm, Bombyx mori.
Agid:
5898632