IntroductionMarek’s disease virus (MDV) is a highly immunosuppressive alphaherpesvirus. However, whether and how MDV exploits extracellular vesicles (EVs) to evade host immunity, particularly the critical type I interferon (IFN-I) response, remains unknown. We hypothesized that MDV reprograms the EV microRNA (miRNA) cargo to facilitate its escape from the IFN-I-mediated antiviral state.MethodsSmall RNA (sRNA) sequencing was conducted to profile and compare the expression patterns of EV miRNAs in chicken embryo fibroblast (DF-1) cells under four conditions: control, MDV infection, chicken interferon-alpha (chIFN-α) treatment and MDV–chIFN-α co-treatment. Integrative bioinformatic analyzes were employed to identify key differentially expressed miRNAs (DEMs) and predict their target genes within the IFN-I signaling network.ResultsMDV infection and chIFN-α treatment induced fundamentally distinct EV miRNA profiles. Strikingly, MDV infection counteracted the specific EV miRNA signature triggered by chIFN-α. We identified 65 key DEMs with the potential to cooperatively target multiple nodes of the IFN-I pathway. Among these, gga-miR-20a-5p and gga-miR-148a-3p were experimentally validated to directly target the 3′ untranslated regions of the key innate immune sensors cGAS and TLR3, respectively, leading to a significant suppression of downstream IFN-I signaling activation.ConclusionThis study identifies dysregulated EV miRNAs during MDV-interferon antagonism, validating their direct targeting of innate immune sensors. These findings provide new insights into viral pathogenesis and pinpoint specific miRNA-target axes as potential avenues for antiviral intervention.

Introduction: Cystic echinococcosis (CE), a chronic disabling parasitic zoonosis, poses a great threat to public health and livestock production and causes huge economic losses globally. The commercial Quil-A-adjuvanted Eg95 vaccine was empirically effective for CE control; however, it is expensive and has side effects and insufficient immunity. Purpose: This study aimed to employ a novel adjuvant consisting of a delivery system and an immune potentiator and assess its adjuvanticity to Eg95 antigen, thereby developing a safe and cost-effective novel vaccine against the disease. Methods: A ferritin-based Eg95 nanoparticle antigen was prepared and then mixed with a plasmid containing the TLR9 agonist CpG to formulate a novel nanovaccine. The safety and efficacy of the vaccine were evaluated in vitro and in vivo. Results: The nanovaccine induced potent and enduring Eg95-specific humoral and cellular immune responses, as well as protective immunity-associated Th1 polarization supported by the higher ratios of IgG2a/IgG1 and IFN-gamma/IL-4. Meanwhile, this nanovaccines exhibited favorable safety and economic profiles. Conclusion: Our data demonstrated that the ferritin-CpG hybrid is a promising combination adjuvant to upgrade the traditional Quil-A and this combination adjuvant-based nanovaccine presents good potential as an alternative to the commercial one for practical CE control.

Background Spodoptera litura is a harmful pest that feeds on more than 80 species of plants, and can be infected and killed by Spodoptera litura nucleopolyhedrovirus (SpltNPV). SpltNPV-C3 is a type C SpltNPV clone, that was observed and collected in Japan. Compared with type A or type B SpltNPVs, SpltNPV-C3 can cause the rapid mortality of S. litura larvae. Methods In this study, occlusion bodies (OBs) and occlusion-derived viruses (ODVs) of SpltNPV-C3 were purified, and OBs were observed by scanning electron microscopy (SEM). ODVs were observed under a transmission electron microscope (TEM). Results Both OBs and ODVs exhibit morphological characteristics typical of nucleopolyhedroviruses (NPVs).The genome of SpltNPV-C3 was sequenced and analyzed; the total length was 148,634 bp (GenBank accession 780,426,which was submitted as SpltNPV-II), with a G + C content of 45%. A total of 149 predicted ORFs were found. A phylogenetic tree of 90 baculoviruses was constructed based on core baculovirus genes. LC‒MS/MS was used to analyze the proteins of SpltNPV-C3; 34 proteins were found in the purified ODVs, 15 of which were core proteins. The structure of the complexes formed by per os infectivity factors 1, 2, 3 and 4 (PIF-1, PIF-2, PIF-3 and PIF-4) was predicted with the help of the AlphaFold multimer tool and predicted conserved sequences in PIF-3. SpltNPV-C3 is a valuable species because of its virulence, and the analysis of its genome and proteins in this research will be beneficial for pest control efforts.

Ten new drimane meroterpenoids talarines A-J (1-10), along with six known analogues (11-16), were isolated from desert soil-derived fungus Talaromyces pinophilus LD-7. Their 2D structures were elucidated by comprehensive interpretation of NMR and HRESIMS data. Electronic circular dichroism calculation was used to establish their absolute configurations. Compounds 2, 10, and 11 showed antiviral activities toward vesicular stomatitis virus with IC50 values of 18, 15, and 23 nM, respectively. The structure-bioactivity relationship indicated that chlorine substitution at C-5 contributed greatly to their antiviral activities. Finally, we identified a new halogenase outside the biosynthetic gene cluster, which was responsible for C-5 halogenation of the precursor isocoumarin 17 as a tailoring step in chlorinated meroterpenoids assembly.

Multiplex network clustering can identify the common cluster structure shared by all layers, which is of great significance for downstream research such as personalized recommendations of social relationships and the mining of social media communication behaviors. Traditional multiplex network clustering methods rely only on the topological structure and are suitable for networks with clear structures. However, real-world networks often suffer from sparse connectivity and noisy edges. In addition, the current methods based on prior information only use the obtained consensus prior information as a preprocessing method for multiplex networks, and do not fully utilize and integrate the prior information in the clustering process, resulting in a low accuracy. To solve the problem of insufficient utilization of prior information, we propose a Consensus Subspace Graph Regularization (CSGR) approach for multiplex network clustering, which integrates topological information with the consensus prior information of a network. We first construct the consensus prior matrix of a network by a non-overlapping greedy search method, which represents a subset of the network that consists of nodes with high edge density. Then we construct a graph regularization term, which encodes the consensus prior information, to optimize the generation of a joint low-dimensional representation within the consensus subspace for the entire multiplex network. Finally, we employ the consensus prior matrix to denoise each network layer, thereby yielding a precise low-dimensional representation of each layer for consensus subspace fusion. In addition, comprehensive experiments have demonstrated that CSGR improves the clustering accuracy of real-world networks by an average of 1.56%.

Purpose: COVID-19 is rampant throughout the world, which has caused great damage to human lives and seriously hindered the development of the global economy. Aiming at the treatment of SARS-CoV-2, in this study, we proposed a novel fenobody strategy based on ferritin (Fe) self-assembly technology. Methods: The neutralizing nanobody H11-D4 of SARS-CoV-2 fused to the C-terminus of end-modified human ferritin was expressed in E. coli and silkworm baculovirus expression systems. A large number of nanoparticles were successfully self-assembled in silkworms, while relatively few nanoparticles can be observed in the treated products from E. coli by electron microscopy. Subsequently, the fenobody's expression level and neutralizing activity were then evaluated. Results: The results showed that the IC50 of H11-D4 and fenobody Fe-H11-D4 expressed in E. coli were 171.1 nmol L-1 and 20.87 nmol L-1, respectively. However, the IC50 of Fe-HD11-D4 expressed in silkworms was 1.46 nmol L-1 showing better neutralization activity. Conclusion: Therefore, fenobodies can be well self-assembled in silkworm baculovirus expression system, and ferritin self-assembly technology can effectively improve nanobody neutralization activity.

IntroductionGetah virus (GETV) has become a growing potential threat to the global livestock industry and public health. However, little is known about the viral pathogenesis and immune escape mechanisms, leading to ineffective control measures.MethodsIn this study, the antiviral activity of exogenous interferons (IFNs) was assessed by using western blotting (WB), real-time quantitative PCR (RT-qPCR) and indirect immunofluorescence assay (IFA). The comparative transcriptomics among mock- and GETV-infected (MOI = 0.1) ST cells with or without IFN-γ was performed by RNA-seq, and then the transcriptome profiling of GETV-infected ST cells and key pathways and putative factors involved in inhibitory effect of IFN-γ on GETV replication were analyzed by bioinformatics methods and RT-qPCR.ResultsThe results showed that treatment with IFN-γ could suppress GETV replication, and the inhibitory effect lasted for at least 48 h, while the exogenous IFN-α/ω and IFN-λ3 treatments failed to inhibit the viral infection and early replication in vitro. Furthermore, the blueprint of virus-host interaction was plotted by RNA-seq and RT-qPCR, showing systemic activation of inflammatory, apoptotic, and antiviral pathways in response to GETV infection, indicating viral hijacking and inhibition of innate host immunity such as IFN-I/III responses. Last and most importantly, activation of the JAK-STAT signaling pathway and complement and coagulation cascades may be a primary driver for IFN-γ-mediated inhibition of GETV replication.DiscussionThese findings revealed that GETV possessed the capability of viral immune escape and indicated that IFN-γ aided in the prevention and control of GETV, implying the potential molecular mechanism of suppression of GETV by IFN-γ, all of which warrant emphasis or further clarification.

The Dongsha waters of the South China Sea are the location of thick Mesozoic deposits and many mud volcanoes that suggest hydrocarbon accumulation. Knowledge of the mechanism, fluid source and evolution of these mud volcanoes, however, remains inadequate. Ferromanganese nodules have been sampled from several deep-water mud volcanoes in waters around Southeast Dongsha island and provide a unique opportunity to study the fluid origin and evolution of the mud volcanoes in the area. In this contribution, the petrology, geochemistry and C–O isotopes of fluid inclusions in the nodules have been analyzed. The ferromanganese nodules are composed of distinct black Fe–Mn crust layers and light-yellow nucleus that contain quartz that hosts fluid inclusions. The outermost layers mainly consist of iron manganese minerals with high Mn/Fe ratios (0.9∼1.57), whereas the nucleus are dominated by siliceous debris. Positive Ce anomalies (δCe: 1.45∼2.13) and negative Y anomalies in growth layers indicate an anoxic environment, which is conducive to hydromanganite precipitation. In contrast, negative Ce anomalies (δCe: 0.27∼0.96) and positive Y anomalies in the nucleus suggest oxidized conditions. The absence of obvious Eu anomalies may be related to low-temperature hydrothermal conditions. The nucleus is enriched in δ13C (−2.59∼1.09‰) and weakly depleted in δ18O (−4.94∼0.89‰), whereas the growth layers are strongly depleted both in δ13C (−16.94∼–12.57‰) and δ18O (−23.05∼–17.48‰). These values indicate that carbon in the nucleus is mainly inorganic carbon from volcanism, whereas the carbon in the growth layers is related to the leakage of deep old hydrocarbon. The fluid inclusions in the quartz from the nucleus show records of at least two periods of CO2-rich hydrothermal activity and an origin from deep Mesozoic strata before release by mud volcanism. Given the widespread mud volcanism, the quartz from the nucleus may come from debris carried by mud volcano eruptions; the quartz also experienced diagenesis related with micro-organisms. The growth layers are considered to be hydrogenic and affected by the leakage of mature thermogenic hydrocarbons from deep old strata. These findings provide insights into deep mature hydrocarbon sources and thermogenic activity and improve understanding of mud volcanism and deep oil and gas resources in the southeastern Dongsha waters.

与基因相关的细胞死亡途径统称为细胞程序性死亡,细胞焦亡是一种新近发现的依赖炎性半胱天冬氨酸酶,并且伴随炎症反应的细胞程序性死亡方式.细胞焦亡的生物学特征、发生与调控机制都区别于其他细胞死亡方式.简要概述了细胞焦亡的研究历史,并从非编码RNA、细胞应激、受体蛋白和化学物质4个方面详细介绍了细胞焦亡的影响因素和调控机制,以期明确细胞焦亡在先天免疫中的角色.

Spike (S) protein, a homotrimeric glycoprotein, is the most important antigen target for SARS-CoV-2 vaccines. A complete simulation of the advanced structure of this homotrimer during subunit vaccine development is the most likely method to improve its immunoprotective effects. In this study, preparation strategies for the S protein receptor-binding domain, S1 region, and ectodomain trimer nanoparticles were designed using ferritin nanoparticle self-assembly technology. The Bombyx mori baculovirus expression system was used to prepare three nanoparticle vaccines with high expression levels recorded in silkworms. The results in mice showed that the nanoparticle vaccine prepared using this strategy could induce immune responses when administered via both the subcutaneous administration and oral routes. Given the stability of these ferritin-based nanoparticle vaccines, an easy-to-use and low-cost oral immunization strategy can be employed in vaccine blind areas attributed to shortages of ultralow-temperature equipment and medical resources in underdeveloped areas. Oral vaccines are also promising candidates for limiting the spread of SARS-CoV-2 in domestic and farmed animals, especially in stray and wild animals.

猪细小病毒(Porcine parvovirus,PPV)可引起猪(Sus scrofa domesticus)繁殖障碍,疫苗是预防猪细小病毒病、提高母猪抗病力和繁殖率的有效方法.为高效表达猪细小病毒结构蛋白VP2基因及制备猪细小病毒基因工程亚单位疫苗,根据家蚕密码子频率将PPV-VP2的基因序列进行优化合成,并将此目的基因片段连接到转移载体,通过共转染将目的基因重组到orf1629基因缺损的亲本病毒BmBacmid上,获得重组杆状病毒(Recombinant baculovirus)rBmNPV(PPV-VP2).将重组杆状病毒rBmNPV(PPV-VP2)感染家蚕(Bombyx mori)蚕蛹,并用表达的抗原制作疫苗,采用豚鼠(Cavia porcellus)进行疫苗效力试验.经Western blot和质谱分析、电镜观察表达产物.结果表明,在蚕蛹中成功表达出可自组装成病毒空衣壳颗粒的目的蛋白;表达蛋白产物经血凝反应测定,每克蚕蛹表达的抗原效价相当于200剂以上商用疫苗;用表达产物制备成3种抗原剂量的疫苗免疫豚鼠,均产生抗体反应,其抗体能在体外中和PPV病毒.即使每克蚕蛹制备200头份疫苗免疫豚鼠,其血凝抑制效价(hemagglutination inhibition,HI)也达到了1:128,中和抗体效价达到1:83,不低于市售商品苗免疫产生的效价.本研究为猪细小病毒亚单位空衣壳疫苗提供了低成本生产方法.

As an epizootic causative agent, the Getah virus (GETV) can cause moderate illness in horses, lethal disease in foxes, and reproductive disorders and fetal death in pigs. Due to the wide range of hosts and multiple routes of transmission, GETV has become a growing potential threat to the global livestock industry, and even to public health. More attention and research on GETV are urgently needed. In this study, we successfully isolated a novel GETV strain, named BJ0304, from a commercial live vaccine against porcine reproductive and respiratory syndrome virus (PRRSV) and determined its growth kinetics. Then, genetic and phylogenetic analyses were performed. The results revealed that BJ0304 was clustered into Group III, and it was most related to the GETV-V1 strain based on the complete genome sequence. Furthermore, the pathogenicity of the isolate was assessed and found to be a low virulent strain in mice relative to its closest homolog GETV-V1. Finally, mutation and glycosylation analysis showed that a unique mutation (171 T > I) at one amino acid of E2, which affected the glycosylation of E2, may be associated with viral pathogenicity. In summary, the general characteristic of a novel Group III-classified GETV-BJ0304 isolated from commercial live PRRSV vaccine was defined and then mutation/glycosylation-related potential virulence factor was discussed. This study highlights the complexity of GETV transmission routes in swine and the need for more surveillance on commercial animal vaccines, contributes to the understanding of genetic characterization of clinical isolates, provides possible virulence factors in favor of unveiling the viral pathogenesis, and eventually lays the foundation for the prevention and control of GETV.

Complex network clustering problems have been gained great popularity and widespread researches recently, and plentiful optimization algorithms are aimed at this problem. Among these methods, the optimization methods aiming at multiple objectives can break the limitations (e.g., instability) of those optimizing single objective. However, one shortcoming stands out that these methods cannot balance the exploration and exploitation well. In another sentence, it fails to optimize solutions on the basis of the good solutions obtained so far. Inspired by nature, a new optimized method, named multi-objective discrete moth-flame optimization (DMFO) method is proposed to achieve such a tradeoff. Specifically, we redefine the simple flame generation (SFG) and the spiral flight search (SFS) processes with network topology structure to balance exploration and exploitation. Moreover, we present the DMFO in detail utilizing a Tchebycheff decomposition method with an $l_2$ -norm constraint on the direction vector (2-Tch). Besides that, experiments are taken on both synthetic and real-world networks and the results demonstrate the high efficiency and promises of our DMFO when tackling dividing complex networks.

Peste des Petits Ruminants (PPR) is a highly pathogenic disease that is classified as a World Organization for Animal Health (OIE)-listed disease. PPRV mainly infects small ruminants such as goats and sheep. In view of the global and high pathogenicity of PPRV, in this study, we proposed a novel nanoparticle vaccine strategy based on ferritin (Fe) self-assembly technology. Using Helicobacter pylori (H. pylori) ferritin as an antigen delivery vector, a PPRV hemagglutinin (H) protein was fused with ferritin and then expressed and purified in both Escherichia coli (E. coli) and silkworm baculovirus expression systems. Subsequently, the nanoparticle antigens’ expression level, immunogenicity and protective immune response were evaluated. Our results showed that the PPRV hemagglutinin–ferritin (H-Fe) protein was self-assembled in silkworms, while it was difficult to observe the correctly folded nanoparticle in E. coli. Meanwhile, the expression level of the H-Fe protein was higher than that of the H protein alone. Furthermore, the immunogenicity and protective immune response of H-Fe nanoparticle antigens expressed by silkworms were improved compared with the H antigen alone. Particularly, the protective immune response of H-Fe antigens expressed in E. coli did not change, as opposed to the H antigen, which was probably due to the incomplete nanoparticle structure in E. coli. This study indicated that the use of ferritin nanoparticles as antigen delivery carriers could increase the expression of antigen proteins and improve the immunogenicity and immune effect of antigens.

纳米抗体(nanobody,Nb)作为目前已知的能与目标抗原结合的最小单位抗体,在生物医药、临床研究等方面具有良好的应用前景.根据大肠杆菌密码子偏好性优化合成严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome-coronavirus 2,SARS-CoV-2)中和性纳米抗体H11-D4基因,将其克隆到pET28a表达载体上后,转化至大肠杆菌感受态细胞Rosetta(DE3)进行诱导表达,通过镍柱纯化、质谱分析、Western Blot鉴定H11-D4的表达情况并使用中和试验验证其中和活性.研究结果显示,纳米抗体H11-D4可成功在大肠杆菌中表达,最佳诱导条件为IPTG终浓度1.0mmol·L-1,37℃诱导5h.H11-D4抗体的分子量大小约为17.9kD,与预测值相符.经镍柱纯化后,产量为25.16 mg·L-1.透析复性后利用TritonX-114快速有效地去除了内毒素,中和试验成功验证了 H11-D4的中和活性(IC50)为171.1 nmol·L-1,研究结果可为纳米抗体的原核表达和新型冠状病毒肺炎(corona virus disease 2019,COVID-19)的预防及治疗提供基础数据支撑.

干扰素具有广谱抗病毒、抗肿瘤活性,可增强机体抗病毒能力,已经广泛应用于病毒性疾病的防控.小反刍兽疫病毒是危害山羊、绵羊等小反刍动物常见的病毒之一,该病毒的传播对全球养殖业造成了严重的影响.为了表达具有高效抗病毒活性的羊α干扰素(OviIFN-α),将OviIFN-α基因序列根据家蚕密码子偏好性进行优化合成,构建pVL1393-OviIFN-α转移载体,将转移载体与ORF1629缺失型杆状病毒基因组DNA利用脂质体介导的转染法在BmN细胞中进行重组,重组病毒感染家蚕,在蚕血淋巴液中获取重组蛋白OviIFN-α.利用微量细胞病变抑制法在BHK-VSV*GFP系统中检测到该蛋白表达量可达6.5×105 U/mL左右.小反刍兽疫病毒增殖抑制实验显示,羊α干扰素对小反刍兽疫病毒在BHK细胞中的复制产生了明显的抑制作用,IC50为136.93 U左右.该结果为羊α干扰素用于小反刍兽疫疫情防控提供了重要的理论依据.

Interferon (IFN), a critical antiviral cytokine produced by pathogens-induced cells, plays an important role in host innate immune system. In this study, to investigate the inhibition effect of IFN on avian influenza virus (AIV), Chicken Embryo Fibroblasts (CEFs) was infected by H9N2 AIV. The pre-immune state and transcriptome analysis have been observed and performed. The result showed chicken interferon gamma (chIFN-γ) have the most inhibitory effect on H9N2 virus among three types of chicken interferons (chIFNs). Inhibition of chIFN-γ on H9N2 virus was verified by indirect immunofluorescence, RT-qPCR and western blot. The possible signaling pathways induced by chIFN-γ with or without virus were analyzed by transcriptome. The transcriptome data were compared among H9N2-infected, chIFN-γ-treated, chIFN-γ + H9N2-treated, and Control groups. In summary, RNA-sequencing (RNA-seq) data suggested that H9N2 virus infection resulted in corresponding response of certain defensive, inflammatory and metabolism pathways to the virus replication in CEFs. Furthermore, while CEFs were treated with chIFN-γ, many immune-related signaling pathways in cells are affected and altered. Antiviral genes involved in these immune pathways such as interferon regulatory factors, chemokines, interferon-stimulated genes (ISGs) and transcription factors were significantly up-regulated, and showed significant antiviral responses. Compared with virus infected CEFs alone, pretreatment with IFN induced the expression of antiviral genes and activated related antiviral pathways, inhibited the viral replication as result. Our study provided functional annotations for antiviral genes and the basis for studying the mechanism of chIFN-γ mediated response against H9N2 AIV.

Background Prolonged past exposure to secondhand tobacco smoke (SHS) in never-smokers is associated with abnormal lung function and reduced diffusing capacity suggestive of an associated lung tissue injury and damage. The mechanisms by which past SHS exposure may contribute to lung tissue damage are unknown. Elastin is a major constituent of extracellular matrix in lung parenchyma. Objective To determine whether past exposure to SHS is associated with ongoing lung tissue damage as indicated by elevated elastin degradation products that are linked to lung function. Methods We measured the plasma levels of elastin degradation markers (EDM) from 193 never-smoking flight attendants with history of remote SHS exposure in aircraft cabin and 103 nonsmoking flight attendants or sea-level control participants without such history of cabin SHS exposure, and examined those levels versus their lung function with adjustment for covariates. The cabin SHS exposure was estimated based on airline employment history and years of smoking ban enactment. Results The median [interquartile range] plasma EDM level for all participants was 0.30 [0.24-0.36] ng/mL with a total range of 0.16-0.65 ng/mL. Plasma EDM levels were elevated in those with history of exposure to cabin SHS compared to those not exposed (0.33±0.08 vs. 0.26±0.06 ng/mL; age- and sex-adjusted P<0.001). In those with history of cabin SHS-exposure, higher EDM levels were associated with lower diffusing capacity (parameter estimate (PE) [95%CI]=4.2 [0.4-8.0] %predicted decrease per 0.1 ng/mL increase in EDM; P=0.030). Furthermore, EDM levels were inversely associated with FEV1, FEV1/FVC, and FEF25-75 (PE [95%CI]=5.8 [2.1-9.4], 4.0 [2.2-5.7], and 12.5 [5.8-19.2] %predicted decrease per 0.1 ng/mL increase in EDM, respectively; P<0.001). Plasma EDM mediated a substantial fraction of the association of SHS with FEV1, FVC, and FEF25-75 (P<0.05). Conclusions Long after past exposure to SHS, there is ongoing elastin degradation beyond what is expected from the aging process, which likely contributes to lower lung function and reduced pulmonary capillary bed as seen in COPD.

Helicoverpa assulta is a pest that causes severe damage to tobacco, pepper and other cash crops. A local strain of HearNPV-BJ (formerly Helicoverpa assulta nucleopolyhedrovirus (HeasNPV-DJ0031)) was isolated from infected H. assulta larvae in Beijing, which had been regarded as a new kind of baculovirus in previous studies. Describing the biological characteristics of the strain, including its external morphology, internal structure and the pathological characteristics of the infection of various cell lines, can provide references for the identification and function of the virus. HearNPV-BJ virion was defined as a single-nucleocapsid nucleopolyhedrovirus by scanning electron microscopy. QB-Ha-E-5 (H. armigera) and BCIRL-Hz-AM1 (H. zea) cell lines were sensitive to HearNPV-BJ. Undoubtedly modern developed sequencing technology further facilitates the increasing understanding of various strains. The whole genome sequence of the HearNPV-BJ was sequenced and analyzed. The HearNPV-BJ isolate genome was 129, 800 bp nucleotides in length with a G + C content of 38.87% and contained 128 open reading frames (ORFs) encoding predicted proteins of 50 or over 50 amino acids, 67 ORFs in the forward orientation and 61 ORFs in the reverse orientation, respectively. The genome shared 99% sequence identity with Helicoverpa armigera nucleopolyhedrovirus C1 strain (HearNPV-C1), and 103 ORFs had very high homology with published HearNPV sequences. Two bro genes and three hrs were found to be dispersed along the HearNPV-BJ genome. Three of the highest homologs, ORFs with HearNPV, were smaller due to the earlier appearance of the stop codon with unknown functions. P6.9 of HearNPV-BJ, a structural protein, is distinctly different from that of Autographa californica nucleopolyhedrovirus (AcMNPV); its homology with the corresponding gene in HearNPV-C1 was 93.58%. HearNPV-BJ contains 38 core genes identified in other baculoviruses, and phylogenetic analysis indicates HearNPV-BJ belongs to Alphabaculovirus Group II, same as HearNPV-C1. The resulting data provide a better understanding of virion structure, gene function and character of infection. By supplementing the whole-genome sequencing data and Kimura-2 model index, there is more evidence to indicate that HearNPV-BJ may be a variant of Helicoverpa armigera nucleopolyhedrovirus, which also deepens our understanding of the virus species demarcation criteria.

杆状病毒表达系统主要宿主有昆虫细胞和家蚕两种类型,已广泛应用于结构生物学的研究和生物制剂尤其是疫苗的研发.近十余年来由于合成生物学的发展,该系统的发展目标已从单一基因操作转向于需多基因协同操作才能完成的任务.重点研究领域涵盖了难以表达蛋白如膜蛋白的表达量改善、多基因协调表达的兴起尤其是重组腺相关病毒(rAAV)的包装研究、杆状病毒作为哺乳动物基因呈递载体应用的优化、高质量完全人源化蛋白表达系统的建立和完善、通过研究杆状病毒和宿主的互作以改善表达产物的质和量等方面,这极大地拓展了该系统作为一个表达系统的研发应用范围.该系统优化的重点在于进行系统、集成式的改良以提高目的产品的质和量,尤其是应重视宿主侧的遗传背景改良以及病毒和宿主互作的研究.