NDLI: Exogenous gene can be integrated into Nosema bombycis genome by mediating with a non-transposon vector

Content Provider	Springer Nature Link
Author	Guo, Rui Cao, Guangli Lu, Yahong Xue, Renyu Kumar, Dhiraj Hu, Xiaolong Gong, Chengliang
Copyright Year	2016
Abstract	Nosema bombycis, a microsporidium, is a pathogen of pebrine disease of silkworms, and its genomic DNA sequences had been determined. Thus far, the research of gene functions of microsporidium including N. bombycis cannot be performed with gain/loss of function. In the present study, we targeted to construct transgenic N. bombycis. Therefore, hemocytes of the infected silkworm were transfected with a non-transposon vector pIZT/V5-His vector in vivo, and the blood, in which the hemocyte with green fluorescence could be observed, was added to the cultured BmN cells. Furthermore, normal BmN cells were infected with germinated N. bombycis, and the infected cells were transfected with pIZT/V5-His. Continuous fluorescence observations exposed that there were N. bombycis with green fluorescence in some N. bombycis-infected cells, and the extracted genome from the purified N. bombycis spore was used as templates. PCR amplification was carried out with a pair of primers for specifically amplifying the green fluorescence protein (GFP) gene; a specific product representing the gfp gene could be amplified. Expression of the GFP protein through Western blotting also demonstrated that the gfp gene was perfectly inserted into the genome of N. bombysis. These results illustrated that exogenous gene can be integrated into N. bombycis genome by mediating with a non-transposon vector. Our research not only offers a strategy for research on gene function of N. bombycis but also provides an important reference for constructing genetically modified microsporidium utilized for biocontrol of pests.
Starting Page	3093
Ending Page	3098
Page Count	6
File Format	PDF
ISSN	09320113
Journal	Parasitology Research
Volume Number	115
Issue Number	8
e-ISSN	14321955
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2016-04-16
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Transgenic Nosema bombycis Bombyx mori pIZT/V5-His vector BmN cells Transfection Fluorescence Medical Microbiology Microbiology Immunology
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Veterinary Parasitology Insect Science

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