1Introduction Embodied Artificial Intelligence(Embodied AI)has recently become a key research focus[1].It emphasizes agents'abilities to perceive,comprehend,and act in physical worlds to complete tasks.Simulation ...1Introduction Embodied Artificial Intelligence(Embodied AI)has recently become a key research focus[1].It emphasizes agents'abilities to perceive,comprehend,and act in physical worlds to complete tasks.Simulation platforms are essential in this area,as they simulate agent behaviors in set environments and tasks,thereby accelerating algorithm validation and optimization.However,constructing such a platform presents several challenges.展开更多
RESTful APIs have been adopted as the standard way of developing web services,allowing for smooth communication between clients and servers.Their simplicity,scalability,and compatibility have made them crucial to mode...RESTful APIs have been adopted as the standard way of developing web services,allowing for smooth communication between clients and servers.Their simplicity,scalability,and compatibility have made them crucial to modern web environments.However,the increased adoption of RESTful APIs has simultaneously exposed these interfaces to significant security threats that jeopardize the availability,confidentiality,and integrity of web services.This survey focuses exclusively on RESTful APIs,providing an in-depth perspective distinct from studies addressing other API types such as GraphQL or SOAP.We highlight concrete threats-such as injection attacks and insecure direct object references(IDOR)-to illustrate the evolving risk landscape.Our work systematically reviews state-of-the-art detection methods,including static code analysis and penetration testing,and proposes a novel taxonomy that categorizes vulnerabilities such as authentication and authorization issues.Unlike existing taxonomies focused on general web or network-level threats,our taxonomy emphasizes API-specific design flaws and operational dependencies,offering a more granular and actionable framework for RESTful API security.By critically assessing current detection methodologies and identifying key research gaps,we offer a structured framework that advances the understanding and mitigation of RESTful API vulnerabilities.Ultimately,this work aims to drive significant advancements in API security,thereby enhancing the resilience of web services against evolving cyber threats.展开更多
[Objective] This study was to clone the GnRH （gonadotropin-releasing hormone）, and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana ...[Objective] This study was to clone the GnRH （gonadotropin-releasing hormone）, and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana by using RT-PCR techniques. It was conducted with bioinformatics analysis and the in situ hybridization histochemistry of its expression products was studied. [Result] The sequence analy- sis showed that the full cDNA sequence was 1 050 bp with the open reading frame of 1 050 bp, and it encoded 349 amino acid residues. The deduced amino sequence included 7 transmembrane regions, and the predicted molecular mass and isoelectric point were 40.6 kD and 9.54, respectively. The cluster analysis showed that the GnRHR from ＇,4. cerana cerana had close relationship to the GnRHR II from other insects. In situ hybridization showed that Bee-GnRHR staining was specifically localized to the brain, intestine, fat body and testis. [Conclusion] The results indicated that the GnRHR provided molecular bond for the reproduction and metabolism for insects, and suggested a functional role for bee-GnRHR signaling in the coupling of reproduction activities and environment conditions.展开更多
[Objective] This study aimed to clarify the correlation between changes of Apis mel ifera and the nectar secretion characteristics of nectariferous plants. [Method] Considering the nectar secretion characteristics of ...[Objective] This study aimed to clarify the correlation between changes of Apis mel ifera and the nectar secretion characteristics of nectariferous plants. [Method] Considering the nectar secretion characteristics of major and auxiliary nec-tariferous plants, six Apis mel ifera colonies were selected for measure the number of eggs, larvae, pupae and adult bees from Jan. to Dec. in 2012; based on that, their annual change curves were also plotted. [Result] The results showed that there were three peaks of the total number of A. mel ifera workers throughout the year：the first occurred on May 15th, with bees developed into an ideal population for col-lecting pomegranate nectar, and the second and third peaks occurred on July 15th and Oct. 15th, respectively, with bees developed into an ideal population for col ect-ing E. ciliate （Thuab） Hyland. [Conclusion] Prevention of Varroa jacobsoni should be carried out with two or more types of acaricides at the late nectar flow stages of the two nectariferous plants（pomegranate and E. ciliate （Thuab） Hyland） when there was a nectar deficiency. Prevention of Tropilaelaps clareae should be timely per-formed with sublimed sulfur in conjunction with acaricides. This study provides a theoretical basis for the high-quality and high-yielding production of honey, as wel as for the product safety.展开更多
[Objective] The aim was to clone triosephosphate isomerase （TPI） gene from Apis mellifera, and predict the properties of TPI protein with bioinformatic meth- ods. [Method] The TPI gene was firstly cloned by in silic...[Objective] The aim was to clone triosephosphate isomerase （TPI） gene from Apis mellifera, and predict the properties of TPI protein with bioinformatic meth- ods. [Method] The TPI gene was firstly cloned by in silico cloning based on the ex- pressed sequence tags （ESTs） from Unigene of NCBI. Some characters of the TPI protein including hydrophobicity or hydrophilicity, isoelectric point （pl） and secondary structure were analyzed and predicted by the tools of bioinformatics. [Result] The TPI gene from A. mellifera was 1 768 bp in full length and it contained a complete ORF which encoded 247 amino acids; the pl of TPI protein was 8.515; the TPI protein was a member of ~13-fold family. [Conclusion] The in silico cloning based on the expressed sequence tags is a efficient method in practice, and this study will provide more references for further study on A. mellifera at molecular level.展开更多
对Parlay APIs协议和SIP协议中与呼叫相关的部分作了对比,首先提出了两者与呼叫相关的核心概念的映射关系,列举了SIP Server工作在两种常用模式下的具体概念映射模型;随后阐述了Parlay CC API与SIP消息之间的映射,并给出实例加以说明;...对Parlay APIs协议和SIP协议中与呼叫相关的部分作了对比,首先提出了两者与呼叫相关的核心概念的映射关系,列举了SIP Server工作在两种常用模式下的具体概念映射模型;随后阐述了Parlay CC API与SIP消息之间的映射,并给出实例加以说明;接着对二者的地址映射问题进行了讨论;最后给出了一个基于SIP的Par-lay CC API构建基本会话的时序关系。展开更多
Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the...Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene （800 ng mL^-1） and supercoiled plasmid DNA （800 ng mL^-1） under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon＇s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.展开更多
Ascosphaera apis spores containing a dark-colored pigment infect honeybee larvae,resulting in a large-scale collapse of the bee colony due to chalkbrood disease.However,little is known about the pigment or whether it ...Ascosphaera apis spores containing a dark-colored pigment infect honeybee larvae,resulting in a large-scale collapse of the bee colony due to chalkbrood disease.However,little is known about the pigment or whether it plays a role in bee infection caused by A.apis.In this study,the pigment was isolated by alkali extraction,acid hydrolysis,and repeated precipitation.Ultraviolet(UV)analysis revealed that the pigment had a color value of 273,a maximum absorption peak at 195 nm,and a high alkaline solubility(7.67%)and acid precipitability.Further chemical structure analysis of the pigment,including elemental composition,Fourier transform infrared(FTIR)spectroscopy,Raman spectroscopy,mass spectrometry,and nuclear magnetic resonance(NMR),proved that it was a eumelanin with a typical indole structure.The molecular formula of melanin is C10H6O4N2,and its molecular weight is 409 Da.Melanin has hydroxyl,carboxyl,amino,and phenolic groups that can potentially chelate to metal ions.Antioxidant function analyses showed that A.apis melanin had a high scavenging activity against superoxide,hydroxyl,and 2,2-diphenyl-1-picrylhydrazyl(DPPH)radicals,and a high reducing ability to Fe3+.Indirect immunofluorescence assay(IFA),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)analyses showed that A.apis melanin was located on the spore wall.The spore wall localization,antioxidant activity,and metal ion chelating properties of fungal melanin have been suggested to contribute to spore pathogenicity.However,further infection experiments showed that melanin-deficient spores did not reduce the mortality of bee larvae,indicating that melanin does not increase the virulence of A.apis spores.This study is the first report on melanin produced by A.apis,providing an important background reference for further study on its role in A.apis.展开更多
To evaluate Apis mellifera adansonii Latreille impact on fruit and seed yields of Gossypium hirsutum L., its foraging and pollinating activities were studied in Ngaoundere for two seasons. Observations were made on 34...To evaluate Apis mellifera adansonii Latreille impact on fruit and seed yields of Gossypium hirsutum L., its foraging and pollinating activities were studied in Ngaoundere for two seasons. Observations were made on 340 flowers each year and divided in three treatments. The treatments included unlimited flowers access by all visitors; bagged flowers to deny all visits and limited visits by Apis mellifera adansonii only. The worker bees seasonal rhythm of activity, its foraging behaviour, its pollination efficiency, the fruiting rate, the number of seeds per fruit and the percentage of normal seeds were evaluated. Results show that this bee foraged G. hirsutum flowers throughout the whole blooming period. This bee species intensely harvested pollen and nectar. The mean foraging speed was 9.41 flowers per rain in 2009 and 8.41 flowers per min in 2010. The fruiting rate, the number of seeds per fruit and the percentage of normal seeds of unprotected flowers were significantly higher than those of flowers protected from insects. Through its pollination efficiency, Apis mellifera adansonii provoked a significant increment of the fruiting rate by 60.84% in 2009 and 36.48% in 2010, as well as the number of seeds per fruit by 94.16% in 2009 and 31.41% in 2010, and the percentage of normal seeds by 94.23% in 2009 and 33.49% in 2010. The installation ofA. m. adansonii colonies close to G. hirsutum fields could be recommended to increase fruit, seed and honey yields, and pollen production as a hive product in the region.展开更多
Apis mellifera filamentous virus(Am FV)is a large DNA virus that is endemic in honeybee colonies.The genome sequence of the Am FV Swiss isolate(Am FV CH–C05)has been reported,but so far very few molecular studies hav...Apis mellifera filamentous virus(Am FV)is a large DNA virus that is endemic in honeybee colonies.The genome sequence of the Am FV Swiss isolate(Am FV CH–C05)has been reported,but so far very few molecular studies have been conducted on this virus.In this study,we isolated and purified Am FV(Am FV CN)from Chinese honeybee(Apis mellifera)colonies and elucidated its genomics and proteomics.Electron microscopy showed ovoid purified virions with dimensions of 300–500×210–285 nm,wrapping a 3165×40 nm filamentous nucleocapsid in three figure-eight loops.Unlike Am FV CH–C05,which was reported to have a circular genome,our data suggest that Am FV CN has a linear genome of approximately 493 kb.A total of 197 ORFs were identified,among which36 putative genes including 18 baculoviral homologs were annotated.The overall nucleotide similarity between the CN and CH–C05 isolates was 96.9%.Several ORFs were newly annotated in Am FV CN,including homologs of per os infectivity factor 4(PIF4)and a putative integrase.Phylogenomic analysis placed Am FVs on a separate branch within the newly proposed virus class Naldaviricetes.Proteomic analysis revealed 47 Am FV virionassociated proteins,of which 14 had over 50%sequence coverage,suggesting that they are likely to be main structural proteins.In addition,all six of the annotated PIFs(PIF-0–5)were identified by proteomics,suggesting that they may function as entry factors in Am FV infection.This study provides fundamental information regarding the molecular biology of Am FV.展开更多
Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2...Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.62322601).
文摘1Introduction Embodied Artificial Intelligence(Embodied AI)has recently become a key research focus[1].It emphasizes agents'abilities to perceive,comprehend,and act in physical worlds to complete tasks.Simulation platforms are essential in this area,as they simulate agent behaviors in set environments and tasks,thereby accelerating algorithm validation and optimization.However,constructing such a platform presents several challenges.
文摘RESTful APIs have been adopted as the standard way of developing web services,allowing for smooth communication between clients and servers.Their simplicity,scalability,and compatibility have made them crucial to modern web environments.However,the increased adoption of RESTful APIs has simultaneously exposed these interfaces to significant security threats that jeopardize the availability,confidentiality,and integrity of web services.This survey focuses exclusively on RESTful APIs,providing an in-depth perspective distinct from studies addressing other API types such as GraphQL or SOAP.We highlight concrete threats-such as injection attacks and insecure direct object references(IDOR)-to illustrate the evolving risk landscape.Our work systematically reviews state-of-the-art detection methods,including static code analysis and penetration testing,and proposes a novel taxonomy that categorizes vulnerabilities such as authentication and authorization issues.Unlike existing taxonomies focused on general web or network-level threats,our taxonomy emphasizes API-specific design flaws and operational dependencies,offering a more granular and actionable framework for RESTful API security.By critically assessing current detection methodologies and identifying key research gaps,we offer a structured framework that advances the understanding and mitigation of RESTful API vulnerabilities.Ultimately,this work aims to drive significant advancements in API security,thereby enhancing the resilience of web services against evolving cyber threats.
基金Supported by the Science and Technology Planning Project of the Education Department of Shaanxi Province(11JK0618)~~
文摘[Objective] This study was to clone the GnRH （gonadotropin-releasing hormone）, and to investigate its expression in Apis cerana cerana. [Method] The cDNA sequence of GnRHR gene was amplified from Apis cerana cerana by using RT-PCR techniques. It was conducted with bioinformatics analysis and the in situ hybridization histochemistry of its expression products was studied. [Result] The sequence analy- sis showed that the full cDNA sequence was 1 050 bp with the open reading frame of 1 050 bp, and it encoded 349 amino acid residues. The deduced amino sequence included 7 transmembrane regions, and the predicted molecular mass and isoelectric point were 40.6 kD and 9.54, respectively. The cluster analysis showed that the GnRHR from ＇,4. cerana cerana had close relationship to the GnRHR II from other insects. In situ hybridization showed that Bee-GnRHR staining was specifically localized to the brain, intestine, fat body and testis. [Conclusion] The results indicated that the GnRHR provided molecular bond for the reproduction and metabolism for insects, and suggested a functional role for bee-GnRHR signaling in the coupling of reproduction activities and environment conditions.
基金Supported by the Key New Products Development Program of Science and Technology Agency of Yunnan Province(2011BB012)~~
文摘[Objective] This study aimed to clarify the correlation between changes of Apis mel ifera and the nectar secretion characteristics of nectariferous plants. [Method] Considering the nectar secretion characteristics of major and auxiliary nec-tariferous plants, six Apis mel ifera colonies were selected for measure the number of eggs, larvae, pupae and adult bees from Jan. to Dec. in 2012; based on that, their annual change curves were also plotted. [Result] The results showed that there were three peaks of the total number of A. mel ifera workers throughout the year：the first occurred on May 15th, with bees developed into an ideal population for col-lecting pomegranate nectar, and the second and third peaks occurred on July 15th and Oct. 15th, respectively, with bees developed into an ideal population for col ect-ing E. ciliate （Thuab） Hyland. [Conclusion] Prevention of Varroa jacobsoni should be carried out with two or more types of acaricides at the late nectar flow stages of the two nectariferous plants（pomegranate and E. ciliate （Thuab） Hyland） when there was a nectar deficiency. Prevention of Tropilaelaps clareae should be timely per-formed with sublimed sulfur in conjunction with acaricides. This study provides a theoretical basis for the high-quality and high-yielding production of honey, as wel as for the product safety.
基金Supported by Scientific Research Fund of Changzhi University (2010111)~~
文摘[Objective] The aim was to clone triosephosphate isomerase （TPI） gene from Apis mellifera, and predict the properties of TPI protein with bioinformatic meth- ods. [Method] The TPI gene was firstly cloned by in silico cloning based on the ex- pressed sequence tags （ESTs） from Unigene of NCBI. Some characters of the TPI protein including hydrophobicity or hydrophilicity, isoelectric point （pl） and secondary structure were analyzed and predicted by the tools of bioinformatics. [Result] The TPI gene from A. mellifera was 1 768 bp in full length and it contained a complete ORF which encoded 247 amino acids; the pl of TPI protein was 8.515; the TPI protein was a member of ~13-fold family. [Conclusion] The in silico cloning based on the expressed sequence tags is a efficient method in practice, and this study will provide more references for further study on A. mellifera at molecular level.
文摘对Parlay APIs协议和SIP协议中与呼叫相关的部分作了对比,首先提出了两者与呼叫相关的核心概念的映射关系,列举了SIP Server工作在两种常用模式下的具体概念映射模型;随后阐述了Parlay CC API与SIP消息之间的映射,并给出实例加以说明;接着对二者的地址映射问题进行了讨论;最后给出了一个基于SIP的Par-lay CC API构建基本会话的时序关系。
基金supported by the National Basic Research Program of China (2007CB109203 and 2009CB118902)
文摘Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE） and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear crylAh gene （800 ng mL^-1） and supercoiled plasmid DNA （800 ng mL^-1） under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon＇s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic crylAh maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.
基金supported by the Science and Technology Project of the Chongqing Municipal Education Commission(No.KJZD-K202100502)the Natural Science Foundation Project of Chongqing(No.cstc2021jcyj-msxm X0422)+4 种基金the Higher Education Teaching Reform Research Project of the Chongqing Municipal Education Commission(Nos.213132 and KJ173061)the Postgraduate Education and Teaching Reform Research Project of Chongqing Normal University(No.xyjg21012)the Creation&Research Team in College and Universities of Chongqing Municipal Education Commission(No.CXQT21013)the College Student Innovation and Entrepreneurship Training Program Project of the Chongqing Municipal Education Commission(No.202110637013)the Natural Science Foundation Project of Chongqing Normal University(No.13XLB009),China。
文摘Ascosphaera apis spores containing a dark-colored pigment infect honeybee larvae,resulting in a large-scale collapse of the bee colony due to chalkbrood disease.However,little is known about the pigment or whether it plays a role in bee infection caused by A.apis.In this study,the pigment was isolated by alkali extraction,acid hydrolysis,and repeated precipitation.Ultraviolet(UV)analysis revealed that the pigment had a color value of 273,a maximum absorption peak at 195 nm,and a high alkaline solubility(7.67%)and acid precipitability.Further chemical structure analysis of the pigment,including elemental composition,Fourier transform infrared(FTIR)spectroscopy,Raman spectroscopy,mass spectrometry,and nuclear magnetic resonance(NMR),proved that it was a eumelanin with a typical indole structure.The molecular formula of melanin is C10H6O4N2,and its molecular weight is 409 Da.Melanin has hydroxyl,carboxyl,amino,and phenolic groups that can potentially chelate to metal ions.Antioxidant function analyses showed that A.apis melanin had a high scavenging activity against superoxide,hydroxyl,and 2,2-diphenyl-1-picrylhydrazyl(DPPH)radicals,and a high reducing ability to Fe3+.Indirect immunofluorescence assay(IFA),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)analyses showed that A.apis melanin was located on the spore wall.The spore wall localization,antioxidant activity,and metal ion chelating properties of fungal melanin have been suggested to contribute to spore pathogenicity.However,further infection experiments showed that melanin-deficient spores did not reduce the mortality of bee larvae,indicating that melanin does not increase the virulence of A.apis spores.This study is the first report on melanin produced by A.apis,providing an important background reference for further study on its role in A.apis.
文摘To evaluate Apis mellifera adansonii Latreille impact on fruit and seed yields of Gossypium hirsutum L., its foraging and pollinating activities were studied in Ngaoundere for two seasons. Observations were made on 340 flowers each year and divided in three treatments. The treatments included unlimited flowers access by all visitors; bagged flowers to deny all visits and limited visits by Apis mellifera adansonii only. The worker bees seasonal rhythm of activity, its foraging behaviour, its pollination efficiency, the fruiting rate, the number of seeds per fruit and the percentage of normal seeds were evaluated. Results show that this bee foraged G. hirsutum flowers throughout the whole blooming period. This bee species intensely harvested pollen and nectar. The mean foraging speed was 9.41 flowers per rain in 2009 and 8.41 flowers per min in 2010. The fruiting rate, the number of seeds per fruit and the percentage of normal seeds of unprotected flowers were significantly higher than those of flowers protected from insects. Through its pollination efficiency, Apis mellifera adansonii provoked a significant increment of the fruiting rate by 60.84% in 2009 and 36.48% in 2010, as well as the number of seeds per fruit by 94.16% in 2009 and 31.41% in 2010, and the percentage of normal seeds by 94.23% in 2009 and 33.49% in 2010. The installation ofA. m. adansonii colonies close to G. hirsutum fields could be recommended to increase fruit, seed and honey yields, and pollen production as a hive product in the region.
基金the Open Research Fund Program of the State Key Laboratory of Virology of China(Grant No.2019IOV004)the key Research Program of Frontier Sciences of Chinese Academy of Sciences(Grant No.QYZDJ-SSW-SMC021)the National Natural Science Foundation of China(Grant No.31900154 and 31572471)。
文摘Apis mellifera filamentous virus(Am FV)is a large DNA virus that is endemic in honeybee colonies.The genome sequence of the Am FV Swiss isolate(Am FV CH–C05)has been reported,but so far very few molecular studies have been conducted on this virus.In this study,we isolated and purified Am FV(Am FV CN)from Chinese honeybee(Apis mellifera)colonies and elucidated its genomics and proteomics.Electron microscopy showed ovoid purified virions with dimensions of 300–500×210–285 nm,wrapping a 3165×40 nm filamentous nucleocapsid in three figure-eight loops.Unlike Am FV CH–C05,which was reported to have a circular genome,our data suggest that Am FV CN has a linear genome of approximately 493 kb.A total of 197 ORFs were identified,among which36 putative genes including 18 baculoviral homologs were annotated.The overall nucleotide similarity between the CN and CH–C05 isolates was 96.9%.Several ORFs were newly annotated in Am FV CN,including homologs of per os infectivity factor 4(PIF4)and a putative integrase.Phylogenomic analysis placed Am FVs on a separate branch within the newly proposed virus class Naldaviricetes.Proteomic analysis revealed 47 Am FV virionassociated proteins,of which 14 had over 50%sequence coverage,suggesting that they are likely to be main structural proteins.In addition,all six of the annotated PIFs(PIF-0–5)were identified by proteomics,suggesting that they may function as entry factors in Am FV infection.This study provides fundamental information regarding the molecular biology of Am FV.
基金Project (No 2007AA10Z324) supported by the National High-Tech Research and Development Program (863) of China
文摘Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.
