1.Immunogenic evaluation of pseudorabies virus gB protein expressed in the baculovirus-insect cell system.
Jin WANG ; Kai WANG ; Ying ZHANG ; Shuzhen TAN ; Shiqi SUN ; Huichen GUO ; Shuanghui YIN ; Jiaqiang NIU
Chinese Journal of Biotechnology 2025;41(7):2694-2706
Pseudorabies (PR) is an infectious disease caused by the pseudorabies virus (PRV), affecting various domesticated and wild animals. Since pigs are the only natural hosts of PRV, PR poses a serious threat to the pig farming industry. Currently, PR is primarily prevented through vaccination with inactivated vaccines or genetically modified attenuated live vaccines. Developing safe and effective genetically engineered vaccines would facilitate the eradication and control of PR. In this study, the PRV vaccine strain Bartha-K61 was used as the reference strain. The gB protein was expressed via the baculovirus-insect cell expression system. Non-denaturing gel electrophoresis confirmed that the gB protein could form a trimeric structure. The purified gB protein was used to immunize mice, and the immune effect was evaluated by a challenge test. The results showed that the gB antigen induced a strong immune response in mice, with the serum-neutralizing antibody titer above 1:70. The lymphocyte stimulation index reached more than 1.29, and the level of (interferon gamma, IFN-γ) release was higher than 100 pg/mL. After immunization, mice were challenged with the virus at a dose of 104 TCID₅₀/mL, 200 μL per mouse, and the clinical protection rate was 100%. Immunohistochemistry, histopathological section, and tissue viral load results showed that the pathological damage and viral load in the gB-immunized group were significantly lower than those in the PBS group. In summary, the gB protein obtained in this study induced strong humoral and cellular immune responses in mice, laying a foundation for developing a recombinant gB protein subunit vaccine.
Animals
;
Mice
;
Baculoviridae/metabolism*
;
Viral Envelope Proteins/biosynthesis*
;
Herpesvirus 1, Suid/genetics*
;
Pseudorabies/immunology*
;
Swine
;
Pseudorabies Vaccines/genetics*
;
Antibodies, Viral/blood*
;
Insecta/cytology*
;
Mice, Inbred BALB C
;
Female
;
Viral Vaccines/immunology*
2.Analyzing the evolution of insect TMED gene and the expression pattern of silkworm TMED gene.
Chunyang WANG ; Yu GUO ; Haiyin LI ; Ping CHEN
Chinese Journal of Biotechnology 2023;39(12):4996-5013
Transmembrane emp24 domain (TMED) gene is closely related to immune response, signal transduction, growth and disease development in mammals. However, only the Drosophila TMED gene has been reported on insects. We identified the TMED family genes of silkworm, Tribolium castaneum, tobacco moth and Italian bee from their genomes, and found that the TMED family gene composition patterns of one α-class, one β-class, one δ-class and several γ-classes arose in the common ancestor of pre-divergent Hymenoptera insects, while the composition of Drosophila TMED family members has evolved in a unique pattern. Insect TMED family γ-class genes have evolved rapidly, diverging into three separate subclasses, TMED6-like, TMED5-like and TMED3-like. The TMED5-like gene was lost in Hymenoptera, duplicated in the ancestors of Lepidoptera and duplicated in Drosophila. Insect TMED protein not only has typical structural characteristics of TMED, but also has obvious signal peptide. There are seven TMED genes in silkworm, distributed in six chromosomes. One of seven is single exon and others are multi-exons. The complete open reading frame (ORF) sequences of seven TMED genes of silkworm were cloned from larval tissues and registered in GenBank database. BmTMED1, BmTMED2 and BmTMED6 were expressed in all stages and tissues of the silkworm, and all genes were expressed in the 4th and 5th instar and silk gland of the silkworm. The present study revealed the composition pattern of TMED family members, their γ class differentiation and their evolutionary history, providing a basis for further studies on TMED genes in silkworm and other insects.
Animals
;
Bombyx/metabolism*
;
Genes, Insect/genetics*
;
Moths/metabolism*
;
Insecta/metabolism*
;
Drosophila
;
Insect Proteins/metabolism*
;
Phylogeny
;
Mammals/genetics*
3.Strategies for exogenous RNA delivery in RNAi-mediated pest management.
Liu'e GONG ; Shumin YING ; Yafen ZHANG ; Jiaoyu WANG ; Guochang SUN
Chinese Journal of Biotechnology 2023;39(2):459-471
Plant diseases and insect pests threaten the safety of crop production greatly. Traditional methods for pest management are challenged by the problems such as environmental pollution, off-target effects, and resistance of pathogens and insects. New biotechnology-based strategies for pest control are expected to be developed. RNA interference (RNAi) is an endogenous process of gene regulation, which has been widely used to study the gene functions in various organisms. In recent years, RNAi-based pest management has received increasing attention. The effective delivery of the exogenous interference RNA into the targets is a key step in RNAi-mediated plant diseases and pest control. Considerable advances were made on the mechanism of RNAi, and various RNA delivery systems were developed for efficient pest control. Here we review the latest advances on mechanisms and influencing factors of RNA delivery, summarize the strategies of exogenous RNA delivery in RNAi-mediated pest control, and highlight the advantages of nanoparticle complexes in dsRNA delivery.
Animals
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RNA Interference
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Pest Control
;
Insecta/genetics*
;
RNA, Double-Stranded
;
Gene Expression Regulation
4.Advances in receptor-mediated resistance mechanisms of Lepidopteran insects to Bacillus thuringiensis toxin.
Leilei LIU ; Peiwen XU ; Kaiyu LIU ; Wei WEI ; Zhongshen CHANG ; Dahui CHENG
Chinese Journal of Biotechnology 2022;38(5):1809-1823
Bacillus thuringiensis is widely used as an insecticide which is safe and environmentally friendly to humans and animals. One of the important insecticidal mechanisms is the binding of Bt toxins to specific toxin receptors in insect midgut and forming a toxin perforation which eventually leads to insect death. The resistance of target pests to Bt toxins is an important factor hampering the long-term effective cultivation of Bt crops and the continuous use of Bt toxins. This review summarizes the mechanism of insect resistance to Bt toxins from the perspective of important Bt toxin receptors in midgut cells of Lepidopteran insects, which may facilitate the in-depth study of Bt resistance mechanism and pest control.
Animals
;
Bacillus thuringiensis/genetics*
;
Bacillus thuringiensis Toxins
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Bacterial Proteins/metabolism*
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Endotoxins/metabolism*
;
Hemolysin Proteins/metabolism*
;
Insecta/metabolism*
;
Insecticide Resistance/genetics*
;
Insecticides/pharmacology*
;
Pest Control, Biological
5.The nucleocapsid protein of rice stripe virus in cell nuclei of vector insect regulates viral replication.
Wan ZHAO ; Junjie ZHU ; Hong LU ; Jiaming ZHU ; Fei JIANG ; Wei WANG ; Lan LUO ; Le KANG ; Feng CUI
Protein & Cell 2022;13(5):360-378
Rice stripe virus (RSV) transmitted by the small brown planthopper causes severe rice yield losses in Asian countries. Although viral nuclear entry promotes viral replication in host cells, whether this phenomenon occurs in vector cells remains unknown. Therefore, in this study, we systematically evaluated the presence and roles of RSV in the nuclei of vector insect cells. We observed that the nucleocapsid protein (NP) and viral genomic RNAs were partially transported into vector cell nuclei by utilizing the importin α nuclear transport system. When blocking NP nuclear localization, cytoplasmic RSV accumulation significantly increased. In the vector cell nuclei, NP bound the transcription factor YY1 and affected its positive regulation to FAIM. Subsequently, decreased FAIM expression triggered an antiviral caspase-dependent apoptotic reaction. Our results reveal that viral nuclear entry induces completely different immune effects in vector and host cells, providing new insights into the balance between viral load and the immunity pressure in vector insects.
Animals
;
Cell Nucleus
;
Hemiptera/metabolism*
;
Insect Vectors/genetics*
;
Insecta
;
Nucleocapsid Proteins/metabolism*
;
Oryza
;
Plant Diseases
;
Tenuivirus/metabolism*
;
Virus Replication
6.Bioactive phenazines from an earwig-associated Streptomyces sp.
Hao HAN ; Zhi-Kai GUO ; Bo ZHANG ; Mei ZHANG ; Jing SHI ; Wei LI ; Rui-Hua JIAO ; Ren-Xiang TAN ; Hui-Ming GE
Chinese Journal of Natural Medicines (English Ed.) 2019;17(6):475-480
Three new phenazine-type compounds, named phenazines SA-SC (1-3), together with four new natural products (4-7), were isolated from the fermentation broth of an earwig-associated Streptomyces sp. NA04227. The structures of these compounds were determined by extensive analyses of NMR, high resolution mass spectroscopic data, as well as single-crystal X-ray diffraction measurement. Sequencing and analysis of the genome data allowed us to identify the gene cluster (spz) and propose a biosynthetic pathway for these phenazine-type compounds. Additionally, compounds 1-5 exhibited moderate inhibitory activity against acetylcholinesterase (AChE), and compound 3 showed antimicrobial activities against Micrococcus luteus.
Animals
;
Anti-Bacterial Agents
;
chemistry
;
metabolism
;
pharmacology
;
Bacterial Proteins
;
genetics
;
metabolism
;
Crystallography, X-Ray
;
Insecta
;
microbiology
;
Magnetic Resonance Spectroscopy
;
Microbial Sensitivity Tests
;
Micrococcus luteus
;
drug effects
;
Molecular Structure
;
Multigene Family
;
Phenazines
;
chemistry
;
metabolism
;
pharmacology
;
Streptomyces
;
chemistry
;
genetics
;
metabolism
7.Molecular characterization and efficacy evaluation of a transgenic corn event for insect resistance and glyphosate tolerance.
Miao-Miao LIU ; Xiao-Jing ZHANG ; Yan GAO ; Zhi-Cheng SHEN ; Chao-Yang LIN
Journal of Zhejiang University. Science. B 2018;19(8):610-619
A transgenic maize event ZD12-6 expressing a Bacillus thuringiensis (Bt) fusion protein Cry1Ab/Cry2Aj and a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein G10 was characterized and evaluated. Southern blot analysis indicated that ZD12-6 is a single copy integration event. The insert site was determined to be at chromosome 1 by border sequence analysis. Expression analyses of Bt fusion protein Cry1Ab/Cry2Aj and the EPSPS protein G10 suggested that they are both expressed stably in different generations. Insect bioassays demonstrated that the transgenic plants are highly resistant to Asian corn borer (Ostrinia furnacalis), cotton boll worm (Helicoverpa armigera), and armyworm (Mythimna separata). This study suggested that ZD12-6 has the potential to be developed into a commercial transgenic line.
3-Phosphoshikimate 1-Carboxyvinyltransferase/metabolism*
;
Animals
;
Bacillus thuringiensis Toxins
;
Bacterial Proteins/metabolism*
;
China
;
Disease Resistance/genetics*
;
Drug Resistance/genetics*
;
Endotoxins/metabolism*
;
Gene Expression Profiling
;
Glycine/chemistry*
;
Hemolysin Proteins/metabolism*
;
Insecta
;
Plant Diseases/prevention & control*
;
Plants, Genetically Modified/genetics*
;
Zea mays/genetics*
;
Glyphosate
8.Research Advances in Baculovirus Occlusion-derived Virions.
Chinese Journal of Virology 2016;32(1):93-100
Baculoviruses are a family of arthropod-specific viruses that produce two morphologically distinct types of virions (budded and occlusion-derived) in their lifecycle. Baculoviruses establish infection in the midgut of their host via the oral route: occlusion-derived virions have pivotal roles in these processes. This review summarizes the basic characteristics of baculoviruses, and discusses the composition and classification of baculovirus occlusion-derived virions. The latter focuses mainly on the evolution and role of multiple occlusion-derived virions in the lifecycle of baculoviruses. These achievements should aid understanding the evolution and infection mechanisms of baculoviruses.
Animals
;
Baculoviridae
;
genetics
;
growth & development
;
physiology
;
Insecta
;
virology
;
Viral Proteins
;
genetics
;
metabolism
;
Virion
;
genetics
;
growth & development
;
physiology
9.Bt transgenic crops for insect-resistance and modification of Bt protein and utilization of stacking strategy.
Chinese Journal of Biotechnology 2015;31(1):53-64
Insecticidal protein genes from Bacillus thuringiensis are currently the most widely used insect-resistant genes. They have been transferred to many crops for breeding and production. Among them, cotton, maize, potato and other insect-resistant crops are commercialized, creating considerable economic benefit. In this review, we summarized advances in identifying functional genes and transgenic crops for insect resistance, compared different strategies for enhancing vigor of insecticidal protein and utilizing gene stacking as well as listing valuable groups of stacked genes. In addition, the methods for multiple gene transformation was discussed.
Animals
;
Bacterial Proteins
;
genetics
;
Crops, Agricultural
;
genetics
;
Endotoxins
;
genetics
;
Hemolysin Proteins
;
genetics
;
Insecta
;
Plants, Genetically Modified
10.The recombinant expression systems for structure determination of eukaryotic membrane proteins.
Yuan HE ; Kan WANG ; Nieng YAN
Protein & Cell 2014;5(9):658-672
Eukaryotic membrane proteins, many of which are key players in various biological processes, constitute more than half of the drug targets and represent important candidates for structural studies. In contrast to their physiological significance, only very limited number of eukaryotic membrane protein structures have been obtained due to the technical challenges in the generation of recombinant proteins. In this review, we examine the major recombinant expression systems for eukaryotic membrane proteins and compare their relative advantages and disadvantages. We also attempted to summarize the recent technical strategies in the advancement of eukaryotic membrane protein purification and crystallization.
Animals
;
Escherichia coli
;
genetics
;
Eukaryotic Cells
;
metabolism
;
Genetic Vectors
;
HEK293 Cells
;
Humans
;
Insecta
;
cytology
;
genetics
;
Membrane Proteins
;
chemistry
;
genetics
;
metabolism
;
Recombinant Proteins
;
chemistry
;
metabolism
;
Yeasts
;
genetics

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