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Apoptotic protease activating factor-1 (Apaf-1) is an adaptor molecule, essential for activating initiator caspase and downstream effector caspases, which directly cause apoptosis. In fruit flies, nematodes, and mammals, Apaf-1 has been extensively studied. However, the structure and function of Apaf-1 in Lepidoptera remain unclear. This study identified a novel Apaf-1 from Spodoptera litura, named Sl-Apaf-1. Sl-Apaf-1 contains three domains: a CARD domain, as well as NOD and WD motifs, and is very similar to mammalian Apaf-1. Interference of Sl-apaf-1 expression in SL-1 cells blocked apoptosis induced by actinomycin D. Overexpression of Sl-apaf-1 significantly enhances apoptosis induced by actinomycin D in Sf9/SL-1/U2OS cells, suggesting that the function of Sl-Apaf-1 is evolutionarily conserved. Furthermore, Sl-Apaf-1 could interact with Sl-caspase-5 (a homologue of mammalian caspase-9) and yielded a binding affinity of 1.37 × 106 M–1 according isothermal titration calorimetry assay. Initiator caspase (procaspase-5) of S. litura could be activated by Sl-Apaf-1 (without WD motif) in vitro, and the activated Sl-caspase-5 could cleave Sl-procaspase-1 (a homologue of caspase-3 in mammals), which directly caused apoptosis. This study demonstrates the key role of Sl-Apaf-1 in the apoptosis pathway, suggesting that the apoptosis pathway in Lepidopteran insects and mammals is conserved.
Haihao Ma; Xiumei Yan; Lin Yan; Jingyan Zhao; Jiping Song; Rong Peng; Yongbo Yang; Jianxin Peng; Kaiyu Liu. Identification and Functional Analysis of Apoptotic Protease Activating Factor-1 (Apaf-1) from Spodoptera litura. Insects 2021, 12, 64 .
AMA StyleHaihao Ma, Xiumei Yan, Lin Yan, Jingyan Zhao, Jiping Song, Rong Peng, Yongbo Yang, Jianxin Peng, Kaiyu Liu. Identification and Functional Analysis of Apoptotic Protease Activating Factor-1 (Apaf-1) from Spodoptera litura. Insects. 2021; 12 (1):64.
Chicago/Turabian StyleHaihao Ma; Xiumei Yan; Lin Yan; Jingyan Zhao; Jiping Song; Rong Peng; Yongbo Yang; Jianxin Peng; Kaiyu Liu. 2021. "Identification and Functional Analysis of Apoptotic Protease Activating Factor-1 (Apaf-1) from Spodoptera litura." Insects 12, no. 1: 64.
It is well known that insect larval midgut cadherin protein serves as a receptor of Bacillus thuringiensis (Bt) crystal Cry1Ac or Cry1Ab toxins, since structural mutations and downregulation of cad gene expression are linked with resistance to Cry1Ac toxin in several lepidopteran insects. However, the role of Spodoptera frugiperda cadherin protein (SfCad) in the mode of action of Bt toxins remains elusive. Here, we investigated whether SfCad is involved in susceptibility to Cry1Ab or Cry1Fa toxins. In vivo, knockout of the SfCad gene by CRISPR/Cas 9 did not increase tolerance to either of these toxins in S. frugiperda larvae. In vitro cytotoxicity assays demonstrated that cultured insect TnHi5 cells expressing GFP-tagged SfCad did not increase susceptibility to activated Cry1Ab or Cry1Fa toxins. In contrast, expression of another well recognized Cry1A receptor in this cell line, the ABCC2 transporter, increased the toxicity of both Cry1Ab and Cry1Fa toxins, suggesting that SfABCC2 functions as a receptor of these toxins. Finally, we showed that the toxin-binding region of SfCad did not bind to activated Cry1Ab, Cry1Ac, nor Cry1Fa. All these results support that SfCad is not involved in the mode of action of Cry1Ab or Cry1Fa toxins in S. frugiperda.
Jianfeng Zhang; Minghui Jin; Yanchao Yang; Leilei Liu; Yongbo Yang; Isabel Gómez; Alejandra Bravo; Mario Soberón; Yutao Xiao; Kaiyu Liu. The Cadherin Protein Is Not Involved in Susceptibility to Bacillus thuringiensis Cry1Ab or Cry1Fa Toxins in Spodoptera frugiperda. Toxins 2020, 12, 375 .
AMA StyleJianfeng Zhang, Minghui Jin, Yanchao Yang, Leilei Liu, Yongbo Yang, Isabel Gómez, Alejandra Bravo, Mario Soberón, Yutao Xiao, Kaiyu Liu. The Cadherin Protein Is Not Involved in Susceptibility to Bacillus thuringiensis Cry1Ab or Cry1Fa Toxins in Spodoptera frugiperda. Toxins. 2020; 12 (6):375.
Chicago/Turabian StyleJianfeng Zhang; Minghui Jin; Yanchao Yang; Leilei Liu; Yongbo Yang; Isabel Gómez; Alejandra Bravo; Mario Soberón; Yutao Xiao; Kaiyu Liu. 2020. "The Cadherin Protein Is Not Involved in Susceptibility to Bacillus thuringiensis Cry1Ab or Cry1Fa Toxins in Spodoptera frugiperda." Toxins 12, no. 6: 375.
Bacillus thuringiensis Cry1Ac toxin binds to midgut proteins, as cadherin (CAD) and ABCC2 transporter, to form pores leading to larval death. In cell lines, co-expression of CAD and ABCC2 enhance Cry1Ac toxicity significantly, but the mechanism remains elusive. Here, we show that the expression of Helicoverpa armigera CAD (HaCAD-GFP) in Hi5 cells induces susceptibility to Cry1Ac and enhanced Cry1Ac toxicity when co-expressed with H. armigera ABCC2 (HaABCC2-GFP), since Cry1Ac toxicity increased 735-fold compared to Hi5 cells expressing HaCAD-GFP alone or 28-fold compared to HaABCC2-GFP alone. In contrast, the expression of the Spodoptera litura CAD (SlCAD-GFP) in Hi5 cells did not induce susceptibility to Cry1Ac nor it potentiated Cry1Ac toxicity with HaABCC2-GFP. To identify the CAD regions involved in the enhancement of Cry1Ac toxicity with ABCC2, the different CAD domains were replaced between SlCAD-GFP and HaCad-GFP proteins, and cytotoxicity assays were performed in Hi5 cells in the absence or presence of HaABCC2-GFP. The HaCAD toxin-binding region (TB), specifically the CAD repeat-11, was necessary to enhance Cry1Ac toxicity with ABCC2. We propose that CAD TB is involved in recruiting Cry1Ac to localize it in a good position for its interaction with the ABCC2, resulting in efficient toxin membrane insertion enhancing Cry1Ac toxicity.
Yuemin Ma; Jianfeng Zhang; Yutao Xiao; Yang; Liu; Rong Peng; Alejandra Bravo; Mario Soberón; Ma; Xiao; Peng; Yanchao Yang; Chenxi Liu; Yongbo Yang; Kaiyu Liu. The Cadherin Cry1Ac Binding-Region is Necessary for the Cooperative Effect with ABCC2 Transporter Enhancing Insecticidal Activity of Bacillus thuringiensis Cry1Ac Toxin. Toxins 2019, 11, 538 .
AMA StyleYuemin Ma, Jianfeng Zhang, Yutao Xiao, Yang, Liu, Rong Peng, Alejandra Bravo, Mario Soberón, Ma, Xiao, Peng, Yanchao Yang, Chenxi Liu, Yongbo Yang, Kaiyu Liu. The Cadherin Cry1Ac Binding-Region is Necessary for the Cooperative Effect with ABCC2 Transporter Enhancing Insecticidal Activity of Bacillus thuringiensis Cry1Ac Toxin. Toxins. 2019; 11 (9):538.
Chicago/Turabian StyleYuemin Ma; Jianfeng Zhang; Yutao Xiao; Yang; Liu; Rong Peng; Alejandra Bravo; Mario Soberón; Ma; Xiao; Peng; Yanchao Yang; Chenxi Liu; Yongbo Yang; Kaiyu Liu. 2019. "The Cadherin Cry1Ac Binding-Region is Necessary for the Cooperative Effect with ABCC2 Transporter Enhancing Insecticidal Activity of Bacillus thuringiensis Cry1Ac Toxin." Toxins 11, no. 9: 538.
Bacillus thuringiensis Cry toxins exert their toxicity by forming membrane pores after binding with larval midgut membrane proteins known as receptors. Spodoptera litura and Spodoptera frugiperda belong to the same genus, but S. litura is tolerant to Cry1Ac, while S. frugiperda is susceptible. The mechanism involved in the differential toxicity of Cry1Ac to these insect species is not understood. Amino acid sequences analysis of ABCC2, a well-recognized Cry1Ac receptor, from both species showed high sequence identity. Hi5 insect cells expressing SfABCC2 from S. frugiperda were 65-fold more susceptible than those expressing the SlABCC2 from S. litura. Substitution of fragments, point mutations and deletions between the ABCC2 of the two species revealed that ABCC2 amino acid Q125 from SfABCC2 or E125 from SlABCC2 was key factor for the differential Cry1Ac toxicity to Hi5 cells expressing these receptors. Consistently with this, cells expressing Helicoverpa armigera HaABCC2Q122-GFP, were more susceptible to Cry1Ac than cells expressing HaABCC2E122-GFP mutant. Q125 or E125 is located in a predicted exposed loop 1 region of ABCC2 indicating that this region could be important for Cry1Ac binding. These findings identified a single amino acid residue located in loop 1 of ABCC2 transporter as responsible for the different levels of susceptibility to Cry1Ac among various lepidopteran species.
Leilei Liu; Zuwen Chen; Yanchao Yang; Yutao Xiao; Chenxi Liu; Yuemin Ma; Mario Soberón; Alejandra Bravo; Yongbo Yang; Kaiyu Liu. A single amino acid polymorphism in ABCC2 loop 1 is responsible for differential toxicity of Bacillus thuringiensis Cry1Ac toxin in different Spodoptera (Noctuidae) species. Insect Biochemistry and Molecular Biology 2018, 100, 59 -65.
AMA StyleLeilei Liu, Zuwen Chen, Yanchao Yang, Yutao Xiao, Chenxi Liu, Yuemin Ma, Mario Soberón, Alejandra Bravo, Yongbo Yang, Kaiyu Liu. A single amino acid polymorphism in ABCC2 loop 1 is responsible for differential toxicity of Bacillus thuringiensis Cry1Ac toxin in different Spodoptera (Noctuidae) species. Insect Biochemistry and Molecular Biology. 2018; 100 ():59-65.
Chicago/Turabian StyleLeilei Liu; Zuwen Chen; Yanchao Yang; Yutao Xiao; Chenxi Liu; Yuemin Ma; Mario Soberón; Alejandra Bravo; Yongbo Yang; Kaiyu Liu. 2018. "A single amino acid polymorphism in ABCC2 loop 1 is responsible for differential toxicity of Bacillus thuringiensis Cry1Ac toxin in different Spodoptera (Noctuidae) species." Insect Biochemistry and Molecular Biology 100, no. : 59-65.
Cry toxins produced by Bacillus thuringiensis (Bt) are insecticidal proteins widely used in insect control. Recently, it was shown that ATP-binding cassette transporter proteins (ABC) such as ABCC2, ABCC3, ABCG1 and ABCA2 are implicated in the insecticidal action of Cry toxins as putative receptors. However, the transcriptional regulators involved in the expression of ABC transporter genes remain unknown. Sequence analysis of promoter regions of ABCC2 gene from Helicoverpa armigera and ABCC3 gene from Spodoptera litura Sl-HP cultured cells, revealed the potential participation of Forkhead box protein A (FOXA), a transcription factor that regulates the expression of genes through remodeling chromatin. To determine if FOXA was involved in regulating expression of ABCC2 and ABCC3 genes, the expression of FOXA, ABCC2 and ABCC3 was compared in Sl-HP cells that are sensitive to Cry1Ac toxin with those in S. frugiperda Sf9 cells that are not sensitive to the toxin. Expression levels of those genes were significantly higher in Sl-HP than in Sf9 cells. Transient expression of FOXA in Sf9 cells activated ABCC2 and ABCC3 transcription, which directly correlated with enhanced Cry1Ac-susceptibility in these cells. Silencing of FOXA gene expression by RNAi in H. armigera larvae resulted in a decreased expression of ABCC2 and ABCC3 without affecting expression of other Cry toxin receptor genes such as alkaline phosphatase, aminopeptidase or cadherin. Silencing of FOXA gene expression also resulted in a Cry1Ac-tolerant phenotype since lower mortality and higher pupation rate were observed in diet containing Cry1Ac protoxin in comparison with the control group. These results demonstrate that FOXA up-regulates expression of the Cry1Ac-toxin receptor ABCC2 and ABCC3 genes, and that lower FOXA expression correlates with tolerance to Cry toxin in cell lines and in lepidopteran larvae.
Jianghuai Li; Yuemin Ma; Wanli Yuan; Yutao Xiao; Chenxi Liu; Jia Wang; Jianxin Peng; Rong Peng; Mario Soberón; Alejandra Bravo; Yongbo Yang; Kaiyu Liu. FOXA transcriptional factor modulates insect susceptibility to Bacillus thuringiensis Cry1Ac toxin by regulating the expression of toxin-receptor ABCC2 and ABCC3 genes. Insect Biochemistry and Molecular Biology 2017, 88, 1 -11.
AMA StyleJianghuai Li, Yuemin Ma, Wanli Yuan, Yutao Xiao, Chenxi Liu, Jia Wang, Jianxin Peng, Rong Peng, Mario Soberón, Alejandra Bravo, Yongbo Yang, Kaiyu Liu. FOXA transcriptional factor modulates insect susceptibility to Bacillus thuringiensis Cry1Ac toxin by regulating the expression of toxin-receptor ABCC2 and ABCC3 genes. Insect Biochemistry and Molecular Biology. 2017; 88 ():1-11.
Chicago/Turabian StyleJianghuai Li; Yuemin Ma; Wanli Yuan; Yutao Xiao; Chenxi Liu; Jia Wang; Jianxin Peng; Rong Peng; Mario Soberón; Alejandra Bravo; Yongbo Yang; Kaiyu Liu. 2017. "FOXA transcriptional factor modulates insect susceptibility to Bacillus thuringiensis Cry1Ac toxin by regulating the expression of toxin-receptor ABCC2 and ABCC3 genes." Insect Biochemistry and Molecular Biology 88, no. : 1-11.
Although many insect cell lines derived from various tissues are available, it is unclear whether endogenous receptors of Bacillus thuringiensis (Bt) crystal toxins are expressed in these cell lines. In the present study, we demonstrated that the ovaries-derived Spodoptera litura Sl-HP cell line was susceptible to activated Cry1Ac although larvae of S. litura are not susceptible to the toxin. Assays of the transcriptome revealed that thirteen ATP-binding cassette transporter genes (ABC) were expressed at different levels in this cell line. Of these, the SlABCC3 shared 52-55% amino acid sequence identity with the known Bt toxin receptor ABCC2. RNAi-mediated knockdown targeting SlABCC3 significantly decreased the susceptibility of Sl-HP cells to activated Cry1Ac. Over-expression of the gene strongly increased the susceptibility of Trichoplusia ni Hi5 cells to the toxin. Not only was SlABCC3 comparable to the heterologously expressed Helicoverpa armigera Hacadherin on the receptor-mediated cytotoxicity of activated Cry1Ac to Hi5 cells, but also SlABCC3 and Hacadherin had a strong synergistic effect on cytotoxicity of activated Cry1Ac. These results suggested that Bt toxin receptors-expressing insect cell lines can be used as an alternative model for evaluating cytotoxicity of Bt toxins and studying their mechanisms of action.
Zuwen Chen; Fei He; Yutao Xiao; Chenxi Liu; Jianghuai Li; Yongbo Yang; Hui Ai; Jianxin Peng; Huazhu Hong; Kaiyu Liu. Endogenous expression of a Bt toxin receptor in the Cry1Ac-susceptible insect cell line and its synergistic effect with cadherin on cytotoxicity of activated Cry1Ac. Insect Biochemistry and Molecular Biology 2015, 59, 1 -17.
AMA StyleZuwen Chen, Fei He, Yutao Xiao, Chenxi Liu, Jianghuai Li, Yongbo Yang, Hui Ai, Jianxin Peng, Huazhu Hong, Kaiyu Liu. Endogenous expression of a Bt toxin receptor in the Cry1Ac-susceptible insect cell line and its synergistic effect with cadherin on cytotoxicity of activated Cry1Ac. Insect Biochemistry and Molecular Biology. 2015; 59 ():1-17.
Chicago/Turabian StyleZuwen Chen; Fei He; Yutao Xiao; Chenxi Liu; Jianghuai Li; Yongbo Yang; Hui Ai; Jianxin Peng; Huazhu Hong; Kaiyu Liu. 2015. "Endogenous expression of a Bt toxin receptor in the Cry1Ac-susceptible insect cell line and its synergistic effect with cadherin on cytotoxicity of activated Cry1Ac." Insect Biochemistry and Molecular Biology 59, no. : 1-17.
Human bocavirus (HBoV), a parvovirus, is suspected to be an etiologic agent of respiratory disease and gastrointestinal disease in humans. All mRNAs of HBoV1 are transcribed from a single promoter. In this study, we constructed EGFP and luciferase reporter gene vectors under the control of the HBoV1 full promoter (nt 1–252) and its mutated variants, respectively. Fluorescence microscopy was used to observe expression activities of the EGFP. Dual-luciferase reporter vectors were employed in order to evaluate critical promoter elements and the effect of NS1 protein on promoter activity. The HBoV1 promoter activity was about 2.2-fold and 1.9-fold higher than that of the CMV promoter in 293 T and HeLa cells, respectively. The putative transcription factor binding region of the promoter was identified to be located between nt 96 and nt 145. Mutations introduced in the CAAT box of the HBoV1 promoter reduced promoter activity by 34%, whereas nucleotide substitutions in the TATA box had no effect on promoter activity. The HBoV1 promoter activities in 293 T and HeLa cells, in the presence of NS1 protein, were 2- to 2.5-fold higher than those in the absence of NS1 protein. The HBoV1 promoter was highly active in 293 T and HeLa cell lines, and the sequence from nt 96 to nt 145 was critical for the activity of HBoV1 promoter. The CAAT box, in contrast to the TATA-box, was important for optimum promoter activity. In addition, the transcriptional activity of this promoter could be trans-activated by the viral nonstructural protein NS1 in these cells.
Jingjing Li; Yongbo Yang; Yanming Dong; Yongshu Li; Yu Huang; Qianhui Yi; Kaiyu Liu; Yi Li. Key elements of the human bocavirus type 1 (HBoV1) promoter and its trans-activation by NS1 protein. Virology Journal 2013, 10, 315 -315.
AMA StyleJingjing Li, Yongbo Yang, Yanming Dong, Yongshu Li, Yu Huang, Qianhui Yi, Kaiyu Liu, Yi Li. Key elements of the human bocavirus type 1 (HBoV1) promoter and its trans-activation by NS1 protein. Virology Journal. 2013; 10 (1):315-315.
Chicago/Turabian StyleJingjing Li; Yongbo Yang; Yanming Dong; Yongshu Li; Yu Huang; Qianhui Yi; Kaiyu Liu; Yi Li. 2013. "Key elements of the human bocavirus type 1 (HBoV1) promoter and its trans-activation by NS1 protein." Virology Journal 10, no. 1: 315-315.
Yeast Atg8 and mammalian microtubule‐associated protein light chain 3 (LC3) are landmark proteins essential for autophagy. Here the lepidopteran Atg8, a homolog of LC3, is characterized. Sequence analysis reveals that Atg8 proteins are highly conserved in lepidopteran species. The abundance of endogeous Atg8 and the ratios of Atg8 conjugation to phosphatidylethanolamine (Atg8‐PE)/Atg8 are different among several lepidopteran cell lines and different tissues of Helicoverpa armigera larvae. Both the density of fluorescent pre‐autophagosomal structures with GFP‐Ha Atg8 and the abundance of Atg6 are positively correlated with levels of Atg8‐PE in different cell lines. The mutant GFP‐Atg8G116A has lost the function in punctual formation, suggesting that G116 is important for autophagy. Exogenous factors have significant influences on the conversion of Atg8 in lepidopteran cells. Bacillus thuringiensis enhances the degradation of Atg8 in Spodoptera litura Sl‐HP cells. Atg8‐PE degrades gradually with extension of amino acid starvation, and bafilomycin A1 can block the decrease through the inhibition of autophagosome fusion with lysosome. Interestingly, high pH is more effective than amino acid starvation in Bombyx mori Bme cells to induce the conversion of BmAtg8 to BmAgt8‐PE. Change of the quality of fetal bovine serum in the culture medium results in alteration of the ratio of Atg8‐PE/Atg8 in some lepidopteran cell lines.
Zhongchao Gai; Xiaojuan Zhang; Mayira Islam; Xia Wang; Aiying Li; Yongbo Yang; Yi Li; Jianxin Peng; Huazhu Hong; Kaiyu Liu. CHARACTERIZATION OF ATG8 IN LEPIDOPTERAN INSECT CELLS. Archives of Insect Biochemistry and Physiology 2013, 84, 57 -77.
AMA StyleZhongchao Gai, Xiaojuan Zhang, Mayira Islam, Xia Wang, Aiying Li, Yongbo Yang, Yi Li, Jianxin Peng, Huazhu Hong, Kaiyu Liu. CHARACTERIZATION OF ATG8 IN LEPIDOPTERAN INSECT CELLS. Archives of Insect Biochemistry and Physiology. 2013; 84 (2):57-77.
Chicago/Turabian StyleZhongchao Gai; Xiaojuan Zhang; Mayira Islam; Xia Wang; Aiying Li; Yongbo Yang; Yi Li; Jianxin Peng; Huazhu Hong; Kaiyu Liu. 2013. "CHARACTERIZATION OF ATG8 IN LEPIDOPTERAN INSECT CELLS." Archives of Insect Biochemistry and Physiology 84, no. 2: 57-77.
Bin Sun; Yongshu Li; Yanming Dong; Kaiyu Liu; Yongbo Yang; Yi Li. [Modulation of cellular transcriptional factors by human bocavirus 1 nonstructural protein NP1]. ACTA MICROBIOLOGICA SINICA 2013, 53, 1 .
AMA StyleBin Sun, Yongshu Li, Yanming Dong, Kaiyu Liu, Yongbo Yang, Yi Li. [Modulation of cellular transcriptional factors by human bocavirus 1 nonstructural protein NP1]. ACTA MICROBIOLOGICA SINICA. 2013; 53 (7):1.
Chicago/Turabian StyleBin Sun; Yongshu Li; Yanming Dong; Kaiyu Liu; Yongbo Yang; Yi Li. 2013. "[Modulation of cellular transcriptional factors by human bocavirus 1 nonstructural protein NP1]." ACTA MICROBIOLOGICA SINICA 53, no. 7: 1.
Human bocavirus type 1 (HBoV1) is a newly identified pathogen associated with human respiratory tract illnesses. Previous studies demonstrated that proteins of HBoV1 failed to cause cell death, which is considered as a possible common feature of bocaviruses. However, our work showed that the NP1 of HBoV1 induced apoptotic cell death in Hela cells in the absence of viral genome replication and expression of other viral proteins. Mitochondria apoptotic pathway was involved in the NP1-induced apoptosis that was confirmed by apoptotic characteristics including morphological changes, DNA fragmentation and caspase activation. We also demonstrated that the cell cycle of NP1-transfected Hela cells was transiently arrested at G2/M phase followed by rapid appearance of apoptosis and that the N terminal domain of NP1 was critical to its nuclear localization and function in apoptosis induction in Hela cells. These findings might provide alternative information for further study of mechanism of HBoV1 pathogenesis.
Bin Sun; Yingyue Cai; Yongshu Li; Jingjing Li; Kaiyu Liu; Yi Li; Yongbo Yang. The nonstructural protein NP1 of human bocavirus 1 induces cell cycle arrest and apoptosis in Hela cells. Virology 2013, 440, 75 -83.
AMA StyleBin Sun, Yingyue Cai, Yongshu Li, Jingjing Li, Kaiyu Liu, Yi Li, Yongbo Yang. The nonstructural protein NP1 of human bocavirus 1 induces cell cycle arrest and apoptosis in Hela cells. Virology. 2013; 440 (1):75-83.
Chicago/Turabian StyleBin Sun; Yingyue Cai; Yongshu Li; Jingjing Li; Kaiyu Liu; Yi Li; Yongbo Yang. 2013. "The nonstructural protein NP1 of human bocavirus 1 induces cell cycle arrest and apoptosis in Hela cells." Virology 440, no. 1: 75-83.
Human bocavirus (HBoV) is a parvovirus associated with respiratory and gastrointestinal diseases in humans. Recombinant baculoviruses have been used widely for many years to transduce transiently and stably a variety of mammalian cell types at high frequencies. In this study, to explore further the use of baculovirus as a gene delivery vehicle for study of transcription and translation mechanism of human bocavirus which lacks susceptible cell culture system, two recombinant baculoviruses were constructed: Bac-BoV-EGFP in which the EGFP gene was under the control of the HBoV1 promoter, and Bac-HBoV1 encompassing the nearly whole HBoV1 genome without both termini. The data demonstrated that efficient gene delivery and expression were observed in numerous mammalian cells transduced by Bac-BoV-EGFP and the transduction rate was much greater than that in plasmid-based transfected cells. The analysis of transcription and translation in Bac-HBoV1 transduced A549 cells showed that two transcripts from NP1 gene were detected by RT-PCR and the NP1 was localized in the nucleus, suggesting that the Bac-HBoV1 recombinant baculovirus delivered efficiently the HBoV1 genome into A549 cells. In summary, this system provides a useful tool for analysis of the transcription and translation of some viruses lacking a virus-cell replication system.
Jingjing Li; Bin Sun; Hu Han; Jinfeng Ouyang; Hanchao Yao; Yongbo Yang; Kaiyu Liu; Yi Li. Application of baculovirus as a delivery vehicle for study of transcription and translation mechanism of parvovirus in non-permissive mammalian cells. Journal of Virological Methods 2012, 183, 1 -7.
AMA StyleJingjing Li, Bin Sun, Hu Han, Jinfeng Ouyang, Hanchao Yao, Yongbo Yang, Kaiyu Liu, Yi Li. Application of baculovirus as a delivery vehicle for study of transcription and translation mechanism of parvovirus in non-permissive mammalian cells. Journal of Virological Methods. 2012; 183 (1):1-7.
Chicago/Turabian StyleJingjing Li; Bin Sun; Hu Han; Jinfeng Ouyang; Hanchao Yao; Yongbo Yang; Kaiyu Liu; Yi Li. 2012. "Application of baculovirus as a delivery vehicle for study of transcription and translation mechanism of parvovirus in non-permissive mammalian cells." Journal of Virological Methods 183, no. 1: 1-7.