Sheng wu gong cheng xue bao = Chinese journal of biotechnology最新文献

{"title":"[Teaching practice of anaerobic digestion microorganisms in Microbiology for Environmental Engineering].","authors":"Shuguang Wang, Xiaoliang Luo, Chang Chen, Aijun Lin","doi":"10.13345/j.cjb.260096","DOIUrl":"10.13345/j.cjb.260096","url":null,"abstract":"<p><p>Anaerobic digestion is a process in which organic matter is converted into methane and carbon dioxide through the catalysis of bacteria and archaea under anaerobic conditions. It is of great significance for the natural material cycle and sustainable development. Currently, textbooks of Microbiology for Environmental Engineering provide little information on the functional microorganisms involved in anaerobic digestion. As a result, students have a limited understanding of the complexity and applications of anaerobic microorganisms. To address this issue, we carried out teaching reforms. In the classroom, the importance of anaerobic digestion was first introduced. Then, with the four stages of hydrolysis, acidogenesis, acetogenesis, and methanogenesis as the basic framework, the types, characteristics, and functions of the key functional microorganisms in each stage were explained. A particular emphasis was placed on difficult points such as the syntrophic relationships among microorganisms. Finally, with the adoption of an application case integrating theory and practice, students' knowledge mastery was improved, and good teaching effects were obtained. This article introduces the teaching contents and implementation, aiming to provide references for the revision of textbooks and classroom teaching reforms in courses such as Microbiology for Environmental Engineering, Microbiology, and Biochemistry in higher education institutions.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3764-3776"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Screening and identification of gBD124-interacting proteins in goat epididymal head epithelial cells].","authors":"Jinrong Liu, Yining Yan, Xinyun Huang, Jinxin Wu, Youshe Ren, Chunxiang Zhang","doi":"10.13345/j.cjb.260114","DOIUrl":"https://doi.org/10.13345/j.cjb.260114","url":null,"abstract":"<p><p>This study used the prepared gBD124 monoclonal antibody to explore the interacting proteins of gBD124 through co-immunoprecipitation (Co-IP) combined with mass spectrometry, aiming to provide a basis for further exploration of the role of gBD124 in goat breeding and reproduction. Mice were immunized with the purified recombinant gBD124 protein, and specific monoclonal antibody-producing cell lines were obtained by fusing spleen cells and myeloma cells through the mouse monoclonal hybridoma technology. The prepared gBD124 monoclonal antibody with a titer of 1.12×10⁵ was used for Co-IP experiments, and the possible target proteins were screened by mass spectrometry. Then, the goat epididymal head epithelial cells were used for immunofluorescence co-localization to screen the proteins interacting with gBD124, and the results were further verified by Co-IP. The results showed that five cell lines with titers ranging from 0.89×10⁴ to 1.12×10⁵ were obtained, and the gBD124 monoclonal antibody with the highest titer of 1.12×10⁵ was used in the subsequent experiments. Two possible proteins, lamin A/C (LMNA) and heat shock protein 90α family class A member 1 (HSP90AA1), interacting with gBD124 were screened from the goat epididymal head tissue proteins by Co-IP combined with mass spectrometry. The immunofluorescence co-localization results showed that gBD124 and HSP90AA1 were co-localized in the cytoplasm, while gBD124 and LMNA were co-localized in the nucleus. The Co-IP complex fixed by gBD124 antibody contained HSP90AA1 but no LMNA, indicating that HSP90AA1 was the interacting protein of gBD124. The expression analysis of gBD124 and HSP90AA1 in the goat epididymal tissue by fluorescence quantitative PCR and Western blotting showed that the expression of gBD124 was higher in the epididymal head 1 than in other epididymal tissue (<i>P</i>0.05). HSP90AA1 also exhibited higher expression in the epididymal head (<i>P</i>0.05). This study obtained the gBD124 monoclonal antibody and identified the interacting protein HSP90AA1 of gBD124, providing a reference for clarifying the novel interacting proteins and potential signaling networks of β-defensin 124 in the male reproductive system.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3550-3563"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Transcriptome dataset comparing the initial development of swollen stem versus non-swollen basal stem (root-stem node) in <i>Brassica</i><i>juncea</i> var. <i>tumida</i>].","authors":"Chuanxing Zhang, Jin Wen, Youling Zhang, Zhuoran Tang, Ting Peng, Shilin Wang, Qinglin Tang","doi":"10.13345/j.cjb.260268","DOIUrl":"https://doi.org/10.13345/j.cjb.260268","url":null,"abstract":"<p><p>The economic value of <i>Brassica juncea</i> var. <i>tumida</i> depends on the yield and quality of its swollen stem (known as Qingcaitou). However, the frequent lodging of the stem base during early swelling causes the plant to grow prostrate, severely limiting mechanical harvesting. This data paper presents a transcriptome dataset constructed primarily from RNA-seq data, supplemented by histological, physiological, biochemical, and molecular validation data, to compare the initial development between the swollen stem and the non-swollen basal stem (root-stem node). The core components of this dataset include RNA-seq raw reads (FASTQ files) for both stem types. The accompanying validation data consist of histological parameters (cross-sectional area, diameter, phloroglucinol-stained area, and cell compactness), physiological indices (lignin and hemicellulose content), and qRT-PCR validation results for key lignin biosynthesis genes (<i>BjuCCoAOMT1</i> and <i>BjuHCT</i>). The data show that while the swollen stem exhibits advantages in morphological traits such as cross-sectional area and diameter, its phloroglucinol-stained area, cell compactness in stained regions, and lignin and hemicellulose content are significantly lower than those in the root-stem node. Moreover, the root-stem node shows significantly upregulated expression of key lignin biosynthesis genes and specifically activated phenylpropanoid and flavonoid biosynthesis pathways. This dataset reveals quantitative differences in lignin deposition and related gene expression between the two stem types and provides a fundamental resource for molecular marker screening, research on lodging resistance mechanisms, and breeding of varieties suitable for mechanical harvesting.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3478-3491"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preperation of an engineered tryptophan-depleted protein in <i>Pichia pastoris</i> and evaluation of its dietary anti-tumor efficacy.","authors":"Huijia Dai, Yuluo Chen, Yaying Xie, Wenhui Wang, Jinxiu Zhou, Jiayi Chen, Yujing Yu, Yao Lin, Yide Huang","doi":"10.13345/j.cjb.260191","DOIUrl":"https://doi.org/10.13345/j.cjb.260191","url":null,"abstract":"<p><p>Targeting amino acid metabolism has emerged as a promising strategy for cancer therapy. To investigate the feasibility of tryptophan restriction as a dietary intervention strategy to suppress tumor growth <i>in vivo</i>, this study developed a novel tryptophan-depleted protein (TDP) through protein engineering combined with the <i>Pichia pastoris</i> expression system, for dietary intervention to suppress tumor growth. Based on the human serum albumin (HSA) scaffold, we designed the TDP by mutating tryptophan at position 238 to phenylalanine (Phe), and successfully expressed it in <i>P</i>. <i>pastoris</i>. Following purification by hollow fiber membrane ultrafiltration and liquid chromatography, High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis confirmed that the tryptophan content in the purified TDP was only 0.35 μg/g. In a gastric adenocarcinoma xenograft model, the TDP-based tryptophan-restricted diet significantly suppressed tumor growth compared with the control group receiving a complete protein diet (<i>P</i>0.001), resulting in a 64% reduction in tumor weight. This study provides a proof-of-concept for developing engineered dietary proteins as foods for special medical purposes (FSMPs), and establishes a novel strategic framework for dietary intervention in adjuvant cancer therapy.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3683-3698"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Immunogenicity of <i>Mycobacterium tuberculosis</i> Rv0394c protein and its diagnosis potential for bovine tuberculosis].","authors":"Zhengzhong Xu, Yan Gao, Changlin Fan, Gaoya Wu, Fei Pan, Nan Wang, Hongjin Zhao, Chengkun Zheng, Xiang Chen, Xin'an Jiao","doi":"10.13345/j.cjb.250896","DOIUrl":"https://doi.org/10.13345/j.cjb.250896","url":null,"abstract":"<p><p>To evaluate the potential of <i>Mycobacterium tuberculosis</i> Rv0394c protein for use in the diagnosis of bovine tuberculosis, the Rv0394c protein was expressed and purified via a prokaryotic expression system. Its immunogenicity was assessed in mouse and guinea pig models, and its stimulatory effect in peripheral blood IFN-γ release assays was analyzed. The gene fragment of Rv0394c was introduced into the prokaryotic expression vector pColdI <i>via</i> homologous recombination to construct the recombinant plasmid pColdI-Rv0394c, which was transformed into <i>Escherichia coli</i> BL21(DE3) for induced expression. The purified recombinant protein was used to immunize mice to evaluate its immunogenicity. Its diagnostic efficacy as a skin hypersensitivity antigen was assessed in guinea pigs sensitized with inactivated <i>Mycobacterium bovis</i>. Additionally, its potential as a specific stimulating antigen was evaluated by the peripheral blood IFN-γ release assay for bovine tuberculosis. The Rv0394c fusion protein with good immunogenicity was successfully constructed and purified. Mouse experiments showed that Rv0394c stimulated splenic lymphocytes from immunized mice to produce high levels of specific IFN-γ. Skin test results demonstrated that Rv0394c elicited strong positive reactions in sensitized guinea pigs. In the bovine peripheral blood IFN-γ release assay, Rv0394c stimulated high levels of IFN-γ production in tuberculosis-positive cattle. When being used in combination with the fusion protein CFP10-ESAT6 (CE), Rv0394c significantly improved the diagnostic sensitivity for bovine tuberculosis. Rv0394c is an immunodominant antigen with the potential to serve as an allergen for skin test diagnosis and as a stimulating antigen for the peripheral blood IFN-γ release assay in bovine tuberculosis, which provides important biological materials for the development of novel diagnostic technologies for bovine tuberculosis.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3519-3527"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[miR-454-3p suppresses the proliferation of abdominal preadipocytes in broilers].","authors":"Chunling Liu, Zixuan Tian, Zheng Wang","doi":"10.13345/j.cjb.250863","DOIUrl":"https://doi.org/10.13345/j.cjb.250863","url":null,"abstract":"<p><p>To elucidate the role and molecular mechanism of miR-454-3p in the development of abdominal fat in broilers, this study analyzed its expression profiles across different tissues and abdominal adipocytes, and evaluated its regulatory effects on proliferation and differentiation of adipocytes. The results showed that miR-454-3p was highly expressed in the heart, breast muscle, brain, and abdominal adipose tissue, but exhibited relatively low expression in the lung, liver, small intestine, and kidney. During cell proliferation, miR-454-3p expression displayed a dynamic pattern characterized by an initial decrease followed by an increase, whereas it gradually declined during adipogenic differentiation. Functional analyses demonstrated that overexpression of miR-454-3p significantly inhibited cell proliferation and lipid droplet accumulation, accompanied by downregulation of proliferation- and adipogenesis-related genes, including <i>CCND1</i>, <i>CDK2</i>, <i>PCNA</i>, <i>C</i>/<i>EBPβ</i>, <i>PPARγ</i>, and <i>PLIN1</i>. Whereas inhibition of miR-454-3p exerted the opposite effects (<i>P</i>0.05). Transcriptomic analysis revealed that genes upregulated by miR-454-3p were enriched in biological processes related to non-adipogenic lineage development and negative regulation of cell proliferation, while downregulated genes were primarily involved in lipid metabolism, adipogenesis, and cell growth-associated pathways. Further validation using bioinformatic prediction, immunoblotting, and dual-luciferase reporter assays confirmed that the transcription factors <i>KLF7</i> and <i>PPARγ</i> are direct targets of miR-454-3p, with conserved binding sites across vertebrates. Rescue experiments further demonstrated that overexpression of <i>KLF7</i> and <i>PPARγ</i> significantly reversed the inhibitory effects of miR-454-3p on preadipocyte proliferation and differentiation (<i>P</i>0.05). In summary, miR-454-3p coordinately suppresses preadipocyte proliferation and differentiation by directly targeting <i>KLF7</i> and <i>PPARγ</i>, providing a theoretical basis for understanding miRNA-mediated post-transcriptional regulation in adipose tissue development.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3564-3579"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813878","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Responsive DNA origami for mRNA delivery].","authors":"Ji Zhao, Jiaming Wang, Jie Song, Pengfei Zhan","doi":"10.13345/j.cjb.260208","DOIUrl":"https://doi.org/10.13345/j.cjb.260208","url":null,"abstract":"<p><p>mRNA technology has emerged as a crucial tool in fields including vaccinology, gene therapy, and regenerative medicine. However, mRNA is susceptible to rapid degradation by ribonucleases (RNases) in physiological environments, and its large molecular weight and negative charge hinder its transmembrane delivery into cells. Existing delivery systems face key bottlenecks such as insufficient serum stability, limited cellular uptake efficiency, and a lack of controlled intracellular release mechanisms. Consequently, there is an urgent need to develop novel delivery platforms that offer both robust protective capabilities and precisely controlled release. In this study, a barrel-shaped DNA nanostructure was constructed using DNA origami technology, and its structural integrity was verified using agarose gel electrophoresis, atomic force microscopy (AFM), and transmission electron microscopy (TEM). The stability of the mRNA-origami complex was further assessed under 10% fetal bovine serum (FBS) conditions, and its cellular uptake behavior was evaluated in HT29 cells. Building on this platform, a stimulus-responsive release strategy was further developed by incorporating an ultraviolet-activatable PC linker, enabling photo-controlled release and thereby achieving responsive mRNA translation expression. The constructed photo-controlled release system allows programmable intracellular dissociation and release upon specific stimuli. By combining carrier structure optimization with a stimulus-responsive mechanism, a comprehensive intelligent mRNA delivery strategy was established. This study provides novel design concepts and experimental paradigms for constructing nucleic acid delivery systems with high stability, efficient cellular uptake, and controllable release capability.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3658-3667"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Screening and characterization of two nanobodies with different CDR3 sequences against porcine CD205].","authors":"Xin Sun, Yangyu Zhang, Yinghui Yang, Zizheng Cai, Yu Lu, Xiuli Feng, Haiwei Cheng","doi":"10.13345/j.cjb.260117","DOIUrl":"https://doi.org/10.13345/j.cjb.260117","url":null,"abstract":"<p><p>The Complementarity-determining region (CDR) is the core region that determines the functions and characteristics of nanobodies. To analyze the impact of the CDR3 region on the developability of nanobodies and antigen-binding activity, this study used two porcine CD205-specific nanobodies (Nb205-1 and Nb205-10) with different CDR3 regions as models and compared them in terms of prokaryotic expression, Ni column purification, and antigen binding, providing references for the screening and evaluation of nanobodies. Phage display technology was employed to screen porcine CD205-specific nanobodies. Positive clones were subjected to sequence alignment, and Swiss-Model was utilized for homology modeling and structural alignment. The <i>Escherichia coli</i> system was employed to evaluate the strain proliferation capacity, <i>in vitro</i> protein expression form, and Ni-NTA affinity purification efficiency. ELISA was used to detect their binding activity with the target protein of porcine CD205 and their antigenic epitopes. Phage-ELISA results demonstrated that two nanobodies, Nb205-1 and Nb205-10, were screened out with high binding activity to the target protein of porcine CD205. Sequence alignment analysis revealed that the framework regions of the two nanobodies were highly conserved, with only several amino acid residue differences in the CDR3 region. Strain Nb205-1 exhibited superior developability, with high protein expression in <i>E</i>. <i>coli</i>, strong binding affinity to Ni-NTA resin, and ease of purification. In contrast, Nb205-10 existed in the inclusion body form, showed weak binding affinity to Ni-NTA resin, and had extremely low purification efficiency. The 3D structural modeling and electrostatic potential analysis revealed that the CDR3 loop of Nb205-1 presented a convex conformation and was rich in positive charges, while Nb205-10 exhibited a relatively flat concave conformation and neutral charge, suggesting that the two may recognize similar antigenic epitopes through different molecular mechanisms. Epitope analysis indicated that the antigenic epitope recognized by Nb205-1 was located in the region ¹MITPLNDWIVAKDCDKTK¹⁸. This study identified two porcine CD205 nanobodies (Nb205-1 and Nb205-10) with similar binding activity, while their developability (expression and purification efficiency) was significantly influenced by sequence differences in the CDR3 region. In future therapeutic nanobody screening, comprehensive evaluation of their developability should be conducted to provide theoretical and practical bases for rational design.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3503-3518"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Construction and characterization of a stable Cas9-expressing monoclonal WSL cell line].","authors":"Meng Yao, Tianlong Li, Bingxiao Sun, Meilin Li, Dingkun Peng, Yijing Wang, Hua-Ji Qiu, Ding Zhang, Lian-Feng Li","doi":"10.13345/j.cjb.250933","DOIUrl":"https://doi.org/10.13345/j.cjb.250933","url":null,"abstract":"<p><p>The molecular mechanism underlying the cellular invasion of African swine fever virus (ASFV) remains incompletely understood, particularly with respect to its key cellular receptors, which constitutes a major bottleneck in the development of effective vaccines and targeted antiviral therapies. To establish a robust experimental platform that supports efficient ASFV replication and enables genome-wide CRISPR screening for systematic identification of host factors involved in viral entry, we employed the CRISPR/Cas9 system to generate stable monoclonal cell lines expressing Cas9 protein based on the wild boar lung (WSL) cell line. Recombinant lentiviruses co-expressing Cas9 and blasticidin resistance gene were packaged <i>via</i> a lentiviral vector system and transduced into WSL cells. Following blasticidin selection, a polyclonal population stably expressing Cas9 was obtained and subjected to fluorescence-activated cell sorting (FACS) to derive monoclonal cell lines. Cas9 expression was determined by Western blotting. To assess the functional gene editing activity of the established clones, we introduced the lentiviruses carrying an <i>EGFP</i> reporter gene along with its specific single-guide RNA (sgRNA) into the monoclonal cell lines, and quantitatively evaluated the editing efficiency <i>via</i> flow cytometry. Furthermore, sgRNAs specifically targeting the ASFV <i>B646L</i> gene and the host <i>TMEM239</i> gene were designed and synthesized to validate the cell line's capacity for editing both viral and host genomic loci. The results demonstrated the successful establishment of seven stable WSL-Cas9 monoclonal cell lines expressing Cas9 protein, among which clone WSL-Cas9-3# exhibited the highest editing efficiency, enabling effective genetic modification of both ASFV and host genes, while maintaining favorable genetic stability and normal growth properties. This study reports the generation of a WSL-Cas9 monoclonal cell line with high CRISPR/Cas9 editing efficiency, stable proliferation, and permissiveness for robust ASFV replication. This engineered cell line provides a reliable platform for future genome-wide functional screening to systematically identify host factors governing ASFV entry and establishes a critical technical foundation for delving into virus-host interactions.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3736-3748"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148814098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"[Expression and immunogenicity of porcine deltacoronavirus S-trimer protein].","authors":"Ruiming Yu, Liping Zhang, Zhongwang Zhang, Peng Zhou, Li Pan, Ligang Yuan, Xinsheng Liu","doi":"10.13345/j.cjb.260066","DOIUrl":"https://doi.org/10.13345/j.cjb.260066","url":null,"abstract":"<p><p>This study aims to express the S-trimer protein of porcine deltacoronavirus (PDCoV) and study its immunogenicity. Targeting the extracellular domain (D20-N1 077) of PDCoV S protein, we introduced the signal peptide sequence (secretoglobin family 1D member 1, SCGB1D1) at the N-terminus and the T4 phage Foldon motif at the C-terminus, and cloned the recombinant sequence into pcDNA3.1(+) vector. The S-trimer fusion protein was expressed in the ExpiCHO expression system and purified, and the molecular weight of the S-trimer fusion protein was analyzed by non-denaturing polyacrylamide gel electrophoresis and molecular sieve gel filtration chromatography. The purified S-trimer fusion protein was used to immunize New Zealand white rabbits to prepare hyperimmune rabbit serum. The titer of specific IgG antibodies in rabbit hyperimmune serum against PDCoV S was measured by indirect ELISA, and the neutralizing antibody titer in the hyperimmune rabbit serum was measured by the serum microneutralization test. The application of hyperimmune rabbit serum was verified by Western blotting and immunofluorescence assay (IFA). The results showed that the PDCoV S-trimer fusion protein was successfully expressed and purified, and its molecular weight was about 660 kDa, which was consistent with the molecular weight of the trimer. Compared with previous studies, this study innovatively introduced the T4 phage Foldon motif into the S protein to stabilize the trimer structure. The S-trimer fusion protein ExpiCHO expressed in the eukaryotic expression system had complete glycosylation and a structure closer to that of the natural S protein. The prepared hyperimmune rabbit serum against rabbit PDCoV S-trimer fusion protein showed the specific antibody titer of 1:218 700 and the neutralizing antibody titer of 1:1 215.9, being able to be applied in laboratory detection of PDCoV by Western blotting and IFA. In summary, the PDCoV S-trimer protein expressed in this study has good immunogenicity, providing biomaterials for the subsequent study of PDCoV S protein structure, the screening of specific monoclonal antibodies, and the development of subunit vaccines.</p>","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"42 8","pages":"3528-3540"},"PeriodicalIF":0.0,"publicationDate":"2026-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}