Design of Self-Integrating Transient Surface Current Density Sensor Integrated Fiber Transmission Link.

The transient surface current density reflects the external coupling of the electromagnetic pulse (EMP) to the tested device. In this paper, the generation mechanism and measurement principle of conductor surface current density are introduced, and the surface current density distribution irradiated by EMP on a typical aircraft structure is simulated and analyzed. The traditional surface current density is usually measured by B-dot antenna, but its output signal is the differential of the measured signal, so additional integrators or numerical integration of the measured data are required. In this paper, a self-integrating surface current sensor based on optical fiber transmission is designed based on the shielded loop antenna with gap structure. The output signal is the real signal waveform to be measured. Compared with coaxial cables, integrated optical fiber transmission improves the anti-interference ability of long-distance transmission signals. At the same time, the design process of the sensor is introduced in detail. The bandwidth of the sensor is 300 kHz~500 MHz, the sensitivity is calibrated at 1.23 (A·m-1)/mV, and the dynamic range is ±25~1400 A·m-1 (35 dB). The surface current of a metal plate is measured in a bounded wave electromagnetic pulse simulator using a detector developed in this paper. The test results show that the developed sensor has good engineering applicability.

A novel system for the generation of baculoviruses mutant for an essential gene.

BACKGROUND: Currently, to delete an essential gene from a baculovirus genome, a cell line stably expressing the gene to be knocked-out should be first generated, which is time-consuming. Alternatively, essential genes can be deleted in E. coli using the λ Red recombination system, which requires an electroporation system. Here, based on homologous recombination in insect cells, we develop an alternative efficient system that requires neither generation of a cell line nor an electroporation system. METHODS AND RESULTS: Using puc19-based inverse PCR, a transfer vector for deleting BmNPV orf92 (Bm92, an essential gene) was efficiently constructed. A copy of Bm92 was introduced into the polyhedrin locus of BmNPV bacmid. The transfer vector was then co-transfected into BmN cell with the modified bacmid to enable homologous recombination at the Bm92 locus. An agarose-free approach was developed for the purification of Bm92-disrupted bacmid viruses in insect cells. Subsequently, BmN cells were co-infected with purified Bm92-disrupted bacmid viruses and unmodified bacmid viruses to allow recombination at the Tn7 insertion site between the two viruses. Finally, bacmid DNA extracted from BmN cells was transformed into chemically-treated competent DH10B cells, and blue colonies containing Bm92-disrupted bacmid were selected using PCR. CONCLUSIONS: For its efficiency and convenience, the system has great potential to be used for the generation of baculovirus knockout mutants.

Discovery and biological evaluation of proteolysis targeting chimeras (PROTACs) as an EGFR degraders based on osimertinib and lenalidomide.

Epidermal growth factor receptor (EGFR) is one of the important and valuable drug targets. Overexpression of EGFR is associated with the development of many types of cancer. In this study, three PROTACs small molecules (16a-16c) were designed, synthesized and evaluated for their cytotoxicity against the growth in different NSCLC cell line and the degradation effect. The bioassay results indicated that 16c has a good inhibition in PC9 cells and H1975 cells, and the corresponding IC50 value was 0.413 µM and 0.657 µM, respectively. Western blotting results demonstrated that compound 16c could serve as an effective EGFRdel19-targeting degrader in PC9 cells.

From aliphatic compounds contaminated soil to active building material: An emerging opportunity for soil remediation and waste utilisation.

Soil contaminated with the production wastewater of 4,4'-diaminostilbene-2,2'-disulfonic acid is extremely hazardous and difficult to bioremediate. In this study, a cost-effective method was developed to reduce the risk of contaminated soil and produce building materials through a combination of ultrasonic processing and solidification/stabilisation. Ultrasonic processing conditions of 5 min at 40 kHz were found to significantly improve the compressive strength of bricks. The results of scanning electron microscopy, X-ray diffraction, and thermogravimetric analysis demonstrated that the enhanced strength was due to the ultrasonic processing controlling the shape and scale of the crystals and microstructure of the cement paste. Furthermore, the effect of the activating agent, CaO, on the leaching toxicity of the bricks was closely related to the curing temperature. Under natural dry conditions (10-25 °C), the leaching toxicity decreased along with the reduction of CaO. However, under high artificial temperature conditions (40 °C), increasing the CaO was beneficial for decreasing the leaching toxicity. The addition of 2.91% CaO was suitable for improving brick performance under both natural dry (10-25 °C) and artificial temperature curing conditions (40 °C). The results of GC-MS revealed that 64.8% and 66.7% of organic species and organic volume, respectively, were reduced in the leachate of the bricks, which was produced by CaO activation and ultrasonic treatments. It was demonstrated that the optimal combined process for cost-effectively transforming hazardous soil to active building materials is feasible.

Uncovering anthocyanin biosynthesis related microRNAs and their target genes by small RNA and degradome sequencing in tuberous roots of sweetpotato.

BACKGROUND: Compared with white-fleshed sweetpotato (WFSP), purple-fleshed sweetpotato (PFSP) is a desirable resource for functional food development because of the abundant anthocyanin accumulation in its tuberous roots. Some studies have shown that the expression regulation mediated by miRNA plays an important role in anthocyanin biosynthesis in plants. However, few miRNAs and their corresponding functions related to anthocyanin biosynthesis in tuberous roots of sweetpotato have been known. RESULTS: In this study, small RNA (sRNA) and degradome libraries from the tuberous roots of WFSP (Xushu-18) and PFSP (Xuzishu-3) were constructed, respectively. Totally, 191 known and 33 novel miRNAs were identified by sRNA sequencing, and 180 target genes cleaved by 115 known ib-miRNAs and 5 novel ib-miRNAs were identified by degradome sequencing. Of these, 121 miRNAs were differently expressed between Xushu-18 and Xuzishu-3. Integrated analysis of sRNA, degradome sequencing, GO, KEGG and qRT-PCR revealed that 26 differentially expressed miRNAs and 36 corresponding targets were potentially involved in the anthocyanin biosynthesis. Of which, an inverse correlation between the expression of ib-miR156 and its target ibSPL in WFSP and PFSP was revealed by both qRT-PCR and sRNA sequencing. Subsequently, ib-miR156 was over-expressed in Arabidopsis. Interestingly, the ib-miR156 over-expressing plants showed suppressed abundance of SPL and a purplish phenotype. Concomitantly, upregulated expression of four anthocyanin pathway genes was detected in transgenic Arabidopsis plants. Finally, a putative ib-miRNA-target model involved in anthocyanin biosynthesis in sweetpotato was proposed. CONCLUSIONS: The results represented a comprehensive expression profiling of miRNAs related to anthocyanin accumulation in sweetpotato and provided important clues for understanding the regulatory network of anthocyanin biosynthesis mediated by miRNA in tuberous crops.

Proteomic-Based Approach to the Proteins Involved in 1-Deoxynojirimycin Accumulation in Silkworm Bombyx mori (Lepidoptera: Bombycidae).

1-Deoxynojirimycin (DNJ) is the most abundant poly-hydroxylated alkaloid in the latex of mulberry leaves and it protects mulberry from insect predation. However, silkworms can survive the poisoning effect of DNJ and accumulate DNJ by consumption of the mulberry leaves. In order to determine the molecular mechanism of DNJ accumulation in silkworm, comparative proteomic analysis was employed to evaluate protein expression in two groups of silkworm bodies (the third instar silkworm bodies had the maximum content of DNJ throughout life, and the newly hatched silkworm bodies had no DNJ). Our results indicated some differentially expressed proteins in the third instar silkworm involved in material metabolism, energy metabolism, oxidation-reduction, detoxification, immune, and transport regulation may correspond to the accumulation of DNJ. Furthermore, the expression levels of five selected differentially expressed protein-encoding genes namely heat shock cognate protein (Hsp 70), glutathione S-transferase sigma 1 (GST), serine protease precursor (Ser), hemolymph protein (30K), and thiol peroxiredoxin (TPx) were investigated by quantitative real-time PCR and the accumulation of DNJ was measured by HPLC. Correlation analysis showed that the expression levels of Hsp70 and Ser were negatively correlated to DNJ accumulation with weak correlation, while 30K, GST, and TPx genes had positive correlation with DNJ accumulation. The findings suggested that these three proteins were probably important in the physiological process of DNJ accumulation in silkworm.

Combination of zero-valent iron and anaerobic microorganisms immobilized in luffa sponge for degrading 1,1,1-trichloroethane and the relevant microbial community analysis.

1,1,1-Trichloroethane (1,1,1-TCA), a dense non-aqueous phase liquid (DNAPL), is relatively slow to remediate naturally; combination of zero-valent iron and immobilized microorganism is a potential means to accelerate DNAPL biodegradation. We first adopted high density luffa sponge (HDLS) as immobilized microorganism carrier. The experimental results demonstrated that (1) the supernatant liquid microorganisms were the optimal immobilized microorganisms for HDLS and (2) the combination of zero-valent iron and immobilized microorganisms accelerated 1,1,1-TCA transformation. Furthermore, in the long-term remediation process, anaerobic microorganisms produced reductant H2S which was beneficial to zero-valent iron PRBs. Through further study of the microbial community, we found that majority of the sulfate-reducing bacteria (SRB) perfectly adapted to the process of 1,1,1-TCA co-metabolism dechlorination. Desulfobulbus and Desulfococcus potentially were the special SRB that contributed significantly to TCA co-metabolism. Additionally, 1,1,1-TCA induced the generation of new SRB and stimulated the growth of majority of dominating methanogens. The results indicated that they played a constructive role in accelerating the dechlorination of 1,1,1-TCA, reduction of sulfate, and improving the production of CH4. Consequently, combination of zero-valent iron and immobilized microorganisms for remediating groundwater by contaminated 1,1,1-TCA is a sustainable and green remediation technology. Especially for groundwater of SO42- type contaminated by 1,1,1-TCA, in the long-term course of combination degradation, cyclic utilization of H2S to prolong the service life of zero-valent iron PRBs. H2 and CH4 generated to capture as potential energy resource. Based on this, a tentative reaction mechanism for Fe0 biodegradation of 1,1,1-TCA was proposed.

High-density natural luffa sponge as anaerobic microorganisms carrier for degrading 1,1,1-TCA in groundwater.

Anaerobic microorganisms were applied to degrade organic contaminants in groundwater with permeable reactive barriers (PRBs). However, anaerobic microorganisms need to select optimal immobilizing material as carrier. The potential of high-density natural luffa sponge (HDLS) (a new variety of luffa) for the immobilization and protection of anaerobic microorganisms was investigated. The HDLS has a dense structure composed of a complicated interwoven fibrous network. Therefore, the abrasion rate of HDLS (0.0068 g s-1) was the smallest among the four carriers [HDLS, ordinary natural luffa sponge (OLS), polyurethane sponge (PS), and gel carrier AQUAPOROUSGEL (APG)]. The results suggest that it also had the greatest water retention (10.26 H2O-g dry carrier-g-1) and SS retention (0.21 g dry carrier-g-1). In comparison to well-established commercialized gel carrier APG, HDLS was of much better mechanical strength, hydrophilicity and stability. Microbial-immobilized HDLS also had the best performance for the remediation of 1,1,1-TCA simulated groundwater. Analysis of the clone libraries from microorganism-immobilized HDLS showed the HDLS could protect microorganisms from the toxicity of 1,1,1-TCA and maintain the stability of microbial community diversity. The mechanism of HDLS immobilizing and protecting microorganisms was proposed as follows. The HDLS had a micron-scale honeycomb structure (30-40 µm) and an irregular ravine structure (4-20 µm), which facilitate the immobilization of anaerobic microorganisms and protect the anaerobic microorganisms.

UV photolysis for enhanced phenol biodegradation in the presence of 2,4,6-trichlorophenol (TCP).

A bacterial strain isolated from activated sludge and identified as Bacillus amyloliquefaciens could biodegrade phenol, but 2,4,6-trichlorophenol (TCP) inhibited phenol biodegradation and biomass growth. UV photolysis converted TCP into dichlorocatechol, monochlorophenol, and dichlorophenol, and this relieved inhibition by TCP. Phenol-removal and biomass-growth rates were significantly accelerated after UV photolysis: the monod maximum specific growth rate (µ(max)) increased by 9% after TCP photolysis, and the half-maximum-rate concentration (K(S)) decreased by 36%. Thus, the major benefit of UV photolysis in this case was to transform TCP into a set of much-less-inhibitory products.

Fungus-insect gall of Phlebopus portentosus.

Phlebopus portentosus is a popular edible wild mushroom found in the tropical Yunnan, China, and northern Thailand. In its natural habitats, a gall often has been found on some plant roots, around which fungal fruiting bodies are produced. The galls are different from common insect galls in that their cavity walls are not made from plant tissue but rather from the hyphae of P. portentosus. Therefore we have termed this phenomenon "fungus-insect gall". Thus far six root mealy bug species in the family Pseudococcidae that form fungus-insect galls with P. portentosus have been identified: Formicococcus polysperes, Geococcus satellitum, Planococcus minor, Pseudococcus cryptus, Paraputo banzigeri and Rastrococcus invadens. Fungus-insect galls were found on the roots of more than 21 plant species, including Delonix regia, Citrus maxima, Coffea arabica and Artocarpus heterophyllus. Greenhouse inoculation trials showed that fungus-insect galls were found on the roots of A. heterophyllus 1 mo after inoculation. The galls were subglobose to globose, fulvous when young and became dark brown at maturation. Each gall harbored one or more mealy bugs and had a chimney-like vent for ventilation and access to the gall. The cavity wall had three layers. Various shaped mealy bug wax deposits were found inside the wall. Fungal hyphae invaded the epidermis of plant roots and sometimes even the cortical cells during the late stage of gall development. The identity of the fungus inside the cavity was confirmed by molecular methods.

Development of an immunochromatographic strip for rapid detection of Pantoea stewartii subsp. stewartii.

A rapid, simple, sensitive, and specific immunochromatographic test strip was developed for the detection of Pantoea stewartii subsp. stewartii (Pss) in corn seed which was soaked overnight and then centrifuged for precipitate re-dissolved as samples. A pair of sensitive monoclonal antibodies for the immunochromatographic test strip was generated by mice immunization and cell fusion. Under optimized conditions, the lower detection limit of the strips for Pss was 1 × 10(5) cfu/mL both in 0.01 M phosphate buffer solution and corn seed samples, with no cross-reactivity with other common plant pathogens. The developed strip is useful and rapid for the detection of Pss in corn seed samples.

How UV photolysis accelerates the biodegradation and mineralization of sulfadiazine (SD).

Sulfadiazine (SD), one of broad-spectrum antibiotics, exhibits limited biodegradation in wastewater treatment due to its chemical structure, which requires initial mono-oxygenation reactions to initiate its biodegradation. Intimately coupling UV photolysis with biodegradation, realized with the internal loop photobiodegradation reactor, accelerated SD biodegradation and mineralization by 35 and 71 %, respectively. The main organic products from photolysis were 2-aminopyrimidine (2-AP), p-aminobenzenesulfonic acid (ABS), and aniline (An), and an SD-photolysis pathway could be identified using C, N, and S balances. Adding An or ABS (but not 2-AP) into the SD solution during biodegradation experiments (no UV photolysis) gave SD removal and mineralization rates similar to intimately coupled photolysis and biodegradation. An SD biodegradation pathway, based on a diverse set of the experimental results, explains how the mineralization of ABS and An (but not 2-AP) provided internal electron carriers that accelerated the initial mono-oxygenation reactions of SD biodegradation. Thus, multiple lines of evidence support that the mechanism by which intimately coupled photolysis and biodegradation accelerated SD removal and mineralization was through producing co-substrates whose oxidation produced electron equivalents that stimulated the initial mono-oxygenation reactions for SD biodegradation.

Depletion of C3orf1/TIMMDC1 inhibits migration and proliferation in 95D lung carcinoma cells.

In our previous study, we identified an association of high expression of c3orf1, also known as TIMMDC1 (translocase of inner mitochondrial membrane domain-containing protein 1), with metastatic characteristics in lung carcinoma cells. To investigate the preliminary function and mechanism of this mitochondrial protein, we depleted C3orf1 expression by introducing siRNA into 95D lung carcinoma cells. We demonstrated that C3orf1 depletion significantly suppressed 95D cell growth and migration. We confirmed C3orf1 localization in the inner mitochondrial membrane and showed that mitochondrial viability, membrane potential, and ATPase activity were remarkably reduced upon depletion of C3orf1. Microarray data indicated that genes involved in regulation of cell death, migration, and cell-cycle arrest were significantly altered after C3orf1 depletion for 48 h. The expression of genes involved in focal adhesion, ECM-receptor interaction, and p53-signaling pathways were notably altered. Furthermore, cell-cycle arrest genes such as CCNG2 and PTEN as well as genes involved in cell migration inhibition, such as TIMP3 and COL3A1, were upregulated after C3orf1 depletion in 95D cells. Concurrently, expression of the migration-promoting gene NUPR1 was markedly reduced, as confirmed by real-time PCR. We conclude that C3orf1 is critical for mitochondrial function, migration, and proliferation in 95D lung carcinoma cells. Depletion of C3orf1 inhibited cell migration and cell proliferation in association with upregulation of genes involved in cell-cycle arrest and cell migration inhibition. These results suggest that C3orf1 (TIMMDC1) may be a viable treatment target for lung carcinoma, and that further study of the role of this protein in lung carcinoma pathogenesis is justified.

Cutting-edge technologies and relevant reaction mechanism difference in treatment of long- and short-chain per- and polyfluoroalkyl substances: A review.

Per- and polyfluoroalkyl substances (PFAS) are emerging contaminants. Compared with short-chain PFAS, long-chain PFAS are more hazardous. Currently, little attention has been paid to the differences in reaction mechanisms between long-chain and short-chain PFAS. This pressing concern has prompted studies about eliminating PFAS and revealing the mechanism difference. The reaction rate and reaction mechanism of each technology was focused on, including (1) adsorption, (2) ion exchange (IX), (3) membrane filtration, (4) advanced oxidation, (5) biotransformation, (6) novel functional material, and (7) other technologies (e.g. ecological remediation, hydrothermal treatment (HT), mechanochemical (MC) technology, micro/nanobubbles enhanced technology, and integrated technologies). The greatest reaction rate k of photocatalysis for long- and short-chain PFAS high up to 63.0 h-1 and 19.7 h-1, respectively. However, adsorption, membrane filtration, and novel functional material remediation were found less suitable or need higher operation demand for treating short-chain PFAS. Ecological remediation is more suitable for treating natural waterbody for its environmentally friendly and fair reaction rate. The other technologies all showed good application potential for both short- and long-chain PFAS, and it was more excellent for long-chain PFAS. The long-chain PFAS can be cleavaged into short-chain PFAS by C-chain broken, -CF2 elimination, nucleophilic substitution of F-, and HF elimination. Furthermore, the application of each type of technology was novelly designed; and suggestions for the future development of PFAS remediation technologies were proposed.

Superb green cycling strategies for microbe-Fe0 neural network-type interaction: Harnessing eight key genes encoding enzymes and mineral transformations to efficiently treat PFOA.

To address time-consuming and efficiency-limited challenges in conventional zero-valent iron (ZVI, Fe0) reduction or biotransformation for perfluorooctanoic acid (PFOA) treatment, two calcium alginate-embedded amendments (biochar-immobilized PFOA-degrading bacteria (CB) and ZVI (CZ)) were developed to construct microbe-Fe0 high-rate interaction systems. Interaction mechanisms and key metabolic pathways were systematically explored using metagenomics and a multi-process coupling model for PFOA under microbe-Fe0 interaction. Compared to Fe0 (0.0076 day-1) or microbe (0.0172 day-1) systems, the PFOA removal rate (0.0426 day-1) increased by 1.5 to 4.6 folds in the batch microbe-Fe0 interaction system. Moreover, Pseudomonas accelerated the transformation of Fe0 into Fe3+, which profoundly impacted PFOA transport and fate. Model results demonstrated microbe-Fe0 interaction improved retardation effect for PFOA in columns, with decreased dispersivity a (0.48 to 0.20 cm), increased reaction rate λ (0.15 to 0.22 h-1), distribution coefficient Kd (0.22 to 0.46 cm3∙g-1), and fraction f´(52 % to 60 %) of first-order kinetic sorption of PFOA in microbe-Fe0 interaction column system. Moreover, intermediates analysis showed that microbe-Fe0 interaction diversified PFOA reaction pathways. Three key metabolic pathways (ko00362, ko00626, ko00361), eight functional genes, and corresponding enzymes for PFOA degradation were identified. These findings provide insights into microbe-Fe0 "neural network-type" interaction by unveiling biotransformation and mineral transformation mechanisms for efficient PFOA treatment.

The molecular binding sequence transformation of soil organic matter and biochar dissolved black carbon antagonizes the transport of 2,4,6-trichlorophenol.

Dissolved organic matter (DOM) and dissolved black carbon (DBC) are significant environmental factors that influence the transport of organic pollutants. However, the mechanisms by which their molecular diversity affects pollutant transport remain unclear. This study elucidates the molecular binding sequence and adsorption sites through which DOM/DBC compounds antagonize the transport of 2,4,6-trichlorophenol (TCP) using column experiments and modelling. DBC exhibits a high TCP adsorption rate (kn = 5.32 × 10-22 mol1-n∙Ln-1∙min-1) and conditional stability constant (logK = 5.19-5.74), indicating a strong binding affinity and antagonistic effect on TCP. This is attributed to the high relative content of lipid/protein compounds in DBC (25.65 % and 30.28 %, respectively). Moreover, the small molecule lipid compounds showed stronger TCP adsorption energy (Ead = -0.0071 eV/-0.0093 eV) in DOM/DBC, combined with two-dimensional correlation spectroscopy model found that DOM/DBC antagonized TCP transport in the environment through binding sequences that transformed from lipid/protein small molecule compounds to lignin/tannin compounds. This study used a multifaceted approach to comprehensively assess the impact of DOM/DBC on TCP transport. It reveals that the molecular diversity of DOM/DBC is a critical factor affecting pollutant transport, providing important insights into the environmental trend and ecological effects of pollutants.

Analysis of the genomic sequence of Philosamia cynthia nucleopolyhedrin virus and comparison with Antheraea pernyi nucleopolyhedrin virus.

BACKGROUND: Two species of wild silkworms, the Chinese oak silkworm (Antheraea pernyi) and the castor silkworm Philosamia cynthia ricini, can acquire a serious disease caused by Nucleopolyhedrin Viruses (NPVs) (known as AnpeNPV and PhcyNPV, respectively). The two viruses have similar polyhedral morphologies and their viral fragments share high sequence similarity. However, the physical maps of the viral genomes and cross-infectivity of the viruses are different. The genome sequences of two AnpeNPV isolates have been published. RESULTS: We sequenced and analyzed the full-length genome of PhcyNPV to compare the gene contents of the two viruses. The genome of PhcyNPV is 125, 376 bp, with a G + C content of 53.65%, and encodes 138 open reading frames (ORFs) of at least 50 amino acids (aa) (GenBank accession number: JX404026). Between PhcyNPV and AnpeMNPV-L and -Z isolates, 126 ORFs are identical, including 30 baculovirus core genes. Nine ORFs were only found in PhcyNPV. Four genes, cath, v-chi, lef 10 and lef 11, were not found in PhcyNPV. However, most of the six genes required for infectivity via the oral route were found in PhcyNPV and in the two AnpeNPV isolates, with high sequence similarities. The pif-3 gene of PhcyNPV contained 59 aa extra amino acids at the N-terminus compared with AnpeNPV. CONCLUSIONS: Most of the genes in PhcyNPV are similar to the two AnpeNPV isolates, including the direction of expression of the ORFs. Only a few genes were missing from PhcyNPV. These data suggest that PhcyNPV and AnpeNPV might be variants of each other, and that the differences in cross-infection might be caused by gene mutations.

Characterization of the open reading frame 7a from Bombyx mori nucleopolyhedrovirus.

The open reading frame 7a of the Bombyx mori nucleopolyhedrovirus (BmNPV orf7a, Bm7a) encodes a predicted polypeptide consisting of 53 amino acid residues. While this polypeptide's homologues were found in certain baculoviruses, its function has not yet been studied. A protein structural assay revealed that the polypeptide expressed from this gene contained a predicted signal peptide at its N-terminus. To investigate the expression of Bm7a, 3'rapid amplification of cDNA ends was used to detect its transcript. A series of recombinant viruses were also constructed to analyze the cellular localization of the Bm7a products. The results indicated that the Bm7a gene is co-transcribed with bv/odv-e26 and orf9. The function of BM7a was studied by using fused enhanced green fluorescence protein (eGFP) as a visual marker to trace the localization of the protein during infection. Fluorescence was observed in the cellular membrane of the infected cells by confocal microscopy. Western blot analysis using an eGFP-antibody also showed that the BM7a-eGFP fusion protein was present in the budded virions (BVs). These data implied that BM7a is a component of the BV. In this work, we identified the function of the Bm7a gene and suggested that it may play a role in BV packaging.

Screening of candidate proteins interacting with IE-2 of Bombyx mori nucleopolyhedrovirus.

IE-2 of Bombyx mori nucleopolyhedrovirus (BmNPV) has been shown to play important roles in baculovirus infection, which are involved in gene expression and viral replication. However, the mechanism remains unknown. In this paper, by TargetP software, four genes, i.e.-2, odv-e26, odv-e56 and BmNPV-gp101 (Ac-orf116) of BmNPV and Autographa californica multiple NPV (AcMNPV) were predicted to be located in mitochondria. By BLAST tool using BmNPV IE-2 protein sequence, 14 NPVs were found to have IE-2 homologues in GenBank, and most of them were predicted to be located in mitochondria, except for that of Antheraea pernyi NPV (AnpeNPV) and Anticarsia gemmatalis NPV (AngeNPV). To observe the subcellular localization of BmNPV IE-2, a recombinant virus overexpressed the IE-2 and eGFP fusion protein was constructed. In infected BmN cells, the fluorescence specifically enriched in the cellular mitochondria. This evidence was accordant with the prediction. Further, Pull-down assay was used to select protein candidates interacting with IE-2 in B. mori cells infected with BmNPV. Of several isolated protein components, sixteen candidates were identified by MALDI-TOF mass-spectrometry, eight baculoviral proteins (ALK-EXO, F protein, IAP-1, LEF-3, LEF-9, ODV-NC42, TLP, and VP39), and eight proteins from B. mori (Actin, ADP/ATP translocase, ATP synthase subunit beta, Beta-tubulin, DNA topoisomerase 2, Histone H4, Soluble guanylyl cyclae alpha-1 subunit, Transketolase). From the functional point of view, most of these proteins were generally divided into two groups, mitochondrial interaction proteins and viral DNA replication proteins. These results implied that the IE-2 had multiple functions involved in regulating viral gene expression, viral replication and also as a component of mitochondrial factors to regulate the cellular energy supply and apoptosis.