Lidija S. Vrhovac , Maria Levkovets , Vladislav Y. Orekhov , Sebastian Westenhoff
{"title":"Refolding of the Deinococcus Radiodurans phytochrome photosensory module and an extended backbone resonance assignment by solution NMR","authors":"Lidija S. Vrhovac , Maria Levkovets , Vladislav Y. Orekhov , Sebastian Westenhoff","doi":"10.1016/j.pep.2025.106699","DOIUrl":"10.1016/j.pep.2025.106699","url":null,"abstract":"<div><div>Solution NMR reveals the structure and dynamics of biomolecules in solution. In particular, the method can detect changes due to perturbation of the molecules, without limiting effects of frozen particles or crystal environments. Phytochromes are photosensors which control the response to red/far-red light in bacteria, fungi and plants, undergo specific structural changes when photoactivated from the Pr to the Pfr state. While structures of phytochromes have been revealed in both states, the structural mechanism of photoconversion remains incompletely understood. Our previous NMR studies of the entire photosensory core module of the <em>D. radiodurans</em> phytochrome have revealed novel structural changes, but the backbone assignment was incomplete. In particular, a lack of the assignment in the protein core hindered more detailed insight in signaling mechanism. Here, we outline an efficient procedure for the refolding of the three-domain, photosensory core fragment of the <em>D. radiodurans</em> phytochrome in its monomeric form. We find that treatment with guanidinium hydrochloride and subsequent dilution effectively refolds the phytochrome, maintaining its functionality. We characterize the refolded protein with solution NMR spectroscopy newly assigning 27 (44) residues in Pr (Pfr), out of which 12 exhibit notable chemical shift perturbation upon photoactivation. The study presents a functional method for purification and refolding of a multidomain protein and opens the door for further structural and dynamic analysis of phytochromes.</div><div><strong>Author summary</strong></div><div>Refolding of proteins is an established method to increase the deuterium-hydrogen exchange of amid bonds in isotopically labeled proteins, which are located deep in the protein core. Yet, the method has to be optimized for each individual protein and in particular for multidomain proteins it is not trivial to find satisfactory experimental conditions. Here we identify a method to refold a <em>D. radiodurans</em> phytochrome construct and characterize the outcome of the procedure using solution NMR and optical spectroscopy. The quick accessibility on whether the refolded phytochrome was functional or not has been obtained from optical spectra, which also made the screening of a number of additives possible. The procedure led to a significant increase in the number of the assigned residues especially in the protein core, close to the photochemically active chromophore, which enables a more detailed investigation of the structure and dynamics throughout the photocycle of the phytochrome.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106699"},"PeriodicalIF":1.4,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143693126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yankang Wang , Hongmei Zhang , Wenjing Shi , Yongheng Rong , Weian Mao , Linhan Wang , Wenzhu Tang , Yun Kong , Shengjun Wang
{"title":"High soluble expression and characterization of human GalNAc transferase T2 and T11 in Escherichia coli","authors":"Yankang Wang , Hongmei Zhang , Wenjing Shi , Yongheng Rong , Weian Mao , Linhan Wang , Wenzhu Tang , Yun Kong , Shengjun Wang","doi":"10.1016/j.pep.2025.106712","DOIUrl":"10.1016/j.pep.2025.106712","url":null,"abstract":"<div><div>The efficient expression of soluble glycosyltransferases from mammalian sources in <em>Escherichia coli</em> (<em>E. coli</em>) remains a significant challenge, often resulting in misfolding and the formation of inclusion bodies. In this study, we investigated strategies to enhance the solubility and catalytic activity of human GalNAc-T2 and GalNAc-T11, two <em>O</em>-glycosyltransferases involved in <em>O</em>-glycosylation of glycoproteins. We found that fusion with maltose-binding protein (MBP) and cellulase catalytic domain (Cel-CD), which led to majority of the fusion proteins being soluble, could increase the solubility of the recombinant proteins. Enzyme activity assays revealed that the fusion glycosyltransferase exhibited significantly higher catalytic efficiency than non-fused enzymes. In addition, the influence of GalNAc-T11 lectin domain on substrate specificity was also determined. The presence of lectin domain had no influence on the recognition of specific substrate and the specific activity of GalNAc-T11. This work offers an efficient approach for the large-scale production of human glycosyltransferases with enhanced bioactivity, highlighting its potential for glycosylation engineering of glycoprotein drugs.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106712"},"PeriodicalIF":1.4,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Impact of arginine addition on protein concentration via ultrafiltration","authors":"Teruo Akuta , Yasunori Kurosawa , Yui Tomioka , Tsutomu Arakawa","doi":"10.1016/j.pep.2025.106701","DOIUrl":"10.1016/j.pep.2025.106701","url":null,"abstract":"<div><div>In this study, we examined the effects of arginine (L-ArgHCl) on ultrafiltration performance, a process with practical significance for not only research but also pharmaceutical applications. Specifically, we assessed the yield and filtration rate of ultrafiltration using rabbit and goat polyclonal antibodies (neutral to basic isoelectric points) as well as model proteins, BSA (acidic) and lysozyme (basic). When a 1 mg/mL protein solution was concentrated approximately 10-fold using a standard commercially available centrifugal ultrafiltration device, the addition of L-ArgHCl significantly improved yield at near-neutral buffer pH. The observed order of improvement was: 20 mM L-ArgHCl >100 mM L-ArgHCl ≈0.5 M NaCl > no addition. A similar trend was observed with BSA, whereas lysozyme achieved slightly higher yields at 100 mM L-ArgHCl. In a 40-fold concentration of rabbit polyclonal antibody from 1 mg/mL, 20 mM L-ArgHCl enhanced yield at pH 6 and 7, but had minimal or no effect at pH 7.5. Notably, at pH 8, high yields were achieved without arginine. L-ArgHCl also accelerated the concentration rate at all pH levels, with greater enhancements observed at higher arginine concentrations. These findings suggest that L-ArgHCl mitigates protein precipitation and solubility loss by reducing protein-protein interactions and nonspecific binding to the ultrafiltration membrane. At pH 8, the increased surface charge of the antibody reduced hydrophobicity, further improving solubility. In summary, the addition of L-ArgHCl, particularly near pH 7, effectively enhanced ultrafiltration performance. This provides a practical strategy for improving protein concentration processes.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106701"},"PeriodicalIF":1.4,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143670737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tianyi Gao , Yun Wang , Tong Zhang , Rou Li , Yue Sun , Kui Zhang , Min Xu , Fei Liu , Boxing Cheng
{"title":"Molecular cloning and functional analysis of a destabilase from Hirudinaria manillensis","authors":"Tianyi Gao , Yun Wang , Tong Zhang , Rou Li , Yue Sun , Kui Zhang , Min Xu , Fei Liu , Boxing Cheng","doi":"10.1016/j.pep.2025.106703","DOIUrl":"10.1016/j.pep.2025.106703","url":null,"abstract":"<div><div>Destabilases are i-type lysozymes with isopeptidase activity and antibacterial and thrombolytic functions. In recent years, destabliases have been identified in an increasing number of invertebrates. <em>Hirudinaria manillensis</em> belonging to the Annelida, as one of the origins of leeches used in traditional Chinese medicine, which has high medicinal value, there have been few reports on the <em>H. manillensis</em> destabliase. In this study, the cDNA sequence of <em>Hmdestabilase</em> was cloned from the salivary glands of <em>H. manillensis</em>. The 3D Structural analysis indicated that Hmdestabilase is similar to other i-type lysozymes in that it adopts an ellipsoidal shape and has a large cleft containing the lysozyme active site. The docking results of Hmdestabilase protein with N-acetylglucosamine trimer molecule have shown that the location and number of hydrogen bonds are one of the key factors for the interaction between the protein and its substrate. The Hmdestabilase fusion protein obtained through the prokaryotic expression system has lysozyme and isopeptidase activities. In addition, Changes in sodium ion concentration in the environment affect the lysozyme activity of Hmdestabilase fusion protein. The above bioinformatic analysis and enzymatic function studies have shown that Hmdestabilase belongs to the i-type lysozyme family. qPCR analysis revealed that blood feeding significantly increased the mRNA expression of <em>Hmdestabilase</em> in the salivary glands of <em>H. manillensis</em>,and successfully priming the innate immune system against harmful microorganisms ingested with food. This study is helpful to elucidate the innate immune response of <em>H. manillensis</em> and promote the artificial breeding of <em>H. manillensis</em>.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"232 ","pages":"Article 106703"},"PeriodicalIF":1.4,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143664386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Preparation and NMR characterization of Aβ peptides at pathological pH","authors":"Xinyue He , Yalan Ma , Naixia Zhang , Chen Zhou","doi":"10.1016/j.pep.2025.106704","DOIUrl":"10.1016/j.pep.2025.106704","url":null,"abstract":"<div><div>Alzheimer's disease (AD) is a neurodegenerative disorder marked by the progressive deterioration of cognitive function. Its pathological hallmarks include the formation of amyloid plaques, which are primarily due to the abnormal aggregation of Aβ peptides. However, the propensity of Aβ peptides for aggregation makes the <em>in vitro</em> preparation very challenging, often resulting in low yield, instability, and impurities. Here in this study, we developed an <em>in vitro</em> method for preparing monomeric Aβ peptides to achieve stable and high-purity samples. Specifically, three strategies including the uses of high concentration of protein denaturant urea, alkaline buffer (ammonium carbonate buffer), and organic solvents (acetonitrile, hexafluoroisopropanol) were applied to prevent Aβ aggregation during the purification. Through an optimized production process, we successfully obtained stable and highly pure <sup>15</sup>N, <sup>13</sup>C-doubly labeled monomeric Aβ40 and Aβ42 peptides suitable for NMR data collection at the pathological acidic pH. Overall, the preparation method presented here offer a robust approach for <em>in vitro</em> production of monomeric Aβ peptides with satisfying purity and reproducibility. Meanwhile, the NMR characterization results for Aβ40 and Aβ42 at pH 6.5 provide useful information for the further biophysical studies involving these two peptides.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106704"},"PeriodicalIF":1.4,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143664387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yuting Ding , Li Jia , Qifubo Geng , Yan Liu , Shaojue Guo , Shuaiying Zhao , Yingying Kong , Quanfang Jin , Guangxu Xu , Jianfeng Xu
{"title":"Screening and functional characterization of nanobodies targeting the transferrin receptor","authors":"Yuting Ding , Li Jia , Qifubo Geng , Yan Liu , Shaojue Guo , Shuaiying Zhao , Yingying Kong , Quanfang Jin , Guangxu Xu , Jianfeng Xu","doi":"10.1016/j.pep.2025.106702","DOIUrl":"10.1016/j.pep.2025.106702","url":null,"abstract":"<div><div>The transferrin receptor (TfR1) mediates the cellular uptake of iron and other molecules, playing a vital role in hematology and tumor growth. Nanobodies (NBs) targeting TfR1 offer promising therapeutic potential due to their small size, high specificity and stability. However, rapid identification of effective nanobodies remains challenging.In this study, the truncated extracellular fragment of human TfR1 was expressed in a prokaryotic system and purified. Immunized camelids provided a source for nanobody libraries, which were screened using phage display and high-throughput strategies to identify candidates with specific TfR1 binding.NB 2D7 with nanomolar-level dissociation constants (KD) were successfully identified.The analysis of Cell Counting Kit-8(CCK8) experiments indicates that the combined treatment of NB2D7 with FeCl<sub>3</sub> can reduce the survival rate of LoVo cells.This research establishes an efficient platform for anti-TfR1 nanobody screening and highlights the therapeutic potential of these nanobodies in cancer treatment and iron metabolism disorders.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106702"},"PeriodicalIF":1.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143634403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M.S.B.W.T.M. Nipuna Sudaraka Tennakoon , Kyoung-Ho Lee , Hyun-Jin Shin
{"title":"Expression of recombinant swine ferritin heavy chain with enhanced solubility in Escherichia coli and simplified purification of ferritin nanoparticles","authors":"M.S.B.W.T.M. Nipuna Sudaraka Tennakoon , Kyoung-Ho Lee , Hyun-Jin Shin","doi":"10.1016/j.pep.2025.106700","DOIUrl":"10.1016/j.pep.2025.106700","url":null,"abstract":"<div><div>Ferritin is a versatile biomolecule used in various medical applications such as drug delivery, vaccines, biological imaging, and diagnostics. The purity and concentration of the ferritin nanoparticles are crucial for achieving excellent outcomes. In this study, we expressed and purified the recombinant swine ferritin heavy chain (rsFTH) as a new candidate for recombinant ferritin nanoparticles. We generated two types of plasmids that can express rsFTH in mammalian and prokaryotic systems. The myc-tagged rsFTH expressed in the mammalian system was purified and ferritin nanoparticles were validated using dynamic light scattering (DLS) and transmission electron microscopy (TEM). A prokaryotic expression system was used to produce rsFTH on a large scale. Protein expression was optimized in Escherichia coli BL21 under varying temperatures and IPTG conditions, and solubility was enhanced by incubation at 25 °C for 18–22 h in auto-induction media, resulting in approximately >50 % protein content in the soluble fraction compared with the pellet. Protein purification was achieved using His-tag affinity chromatography and dialysis with Tris-HCl buffer, yielding adequately pure rsFTH without any apparent protein aggregates. SDS-PAGE and Western blot analysis confirmed the expected molecular weight of rsFTH, and Native-PAGE demonstrated polymerization into higher molecular weight forms. Particle size analysis of purified rsFTH revealed a mean diameter of 15.5 nm, with transmission electron microscopy (TEM) imaging confirming spherical ferritin particles with an iron core. These results suggest that rsFTH can be efficiently expressed and purified in both mammalian and bacterial systems, and has potential applications in nanotechnology and biotechnology.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"231 ","pages":"Article 106700"},"PeriodicalIF":1.4,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143628203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Qiexin Chen , Yao Zhang , Yuxiang Zhang , Xiao Han , Luyao Zhang , Huan Meng , Jian Luo , Rong Yu , Chun Zhang , Yongdong Liu
{"title":"Rational designation and characterization of a novel humanized collagen capable of self-assembling into triple helix and fibrils with D-period","authors":"Qiexin Chen , Yao Zhang , Yuxiang Zhang , Xiao Han , Luyao Zhang , Huan Meng , Jian Luo , Rong Yu , Chun Zhang , Yongdong Liu","doi":"10.1016/j.pep.2025.106698","DOIUrl":"10.1016/j.pep.2025.106698","url":null,"abstract":"<div><div>The triple helix and D-period are distinctive features of native collagen, crucial for its physicochemical properties and bioactivities. However, developing recombinant humanized collagen with D-period features remains elusive. Here, we present a strategy for preparing a novel recombinant humanized collagen using a ‘charged-hydrophobic-charged amino acid’ sequence with the capacity of self-assembling. The hydrophobic amino acids in the middle region are believed to be crucial for the triple helix formation while the charged amino acids at the C- and N-terminal drive the triple-helix to self-assemble into higher-order structures like fibrils, with D-period formation during this process. To prove this concept, the particular fragment of Gly1059-Ala1103 of human type III collagen, featuring arginine (R), lysine (K), aspartic acid (D), and glutamic acid (E)-rich termini and a Glycine-Proline-Alanine (G-P-A) central motif, was selected and repeated to construct a recombinant humanized collagen, designated as rhCL04. This construct successfully formed hierarchical structures, including triple helices, rod-like fibrils, and hydrogels, exhibiting a distinct 10 nm D-period across a broad pH range from 4 to 10. Additionally, cell adhesion and biocompatibility were confirmed using L929 mouse fibroblast cells, demonstrating the ability to promote cell adhesion activity and no significant cytotoxicity. Our study provides valuable insights into the self-assembling mechanisms of native collagens. Moreover, these results highlight the efficacy of this strategy in producing recombinant humanized collagen with collagen-like characteristics. The simplicity and versatility of the approach, combined with the excellent self-assembling properties and biological activity of rhCL04, underscore its potential for biomaterial production.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106698"},"PeriodicalIF":1.4,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143586757","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hao Wang , Xiaohui Zhang , Cunlong Yin , Lu Liu , Liang Chen , Hengfang Tang , Bo Wu , Junfeng Wang
{"title":"High-purity preparation and biophysical characterization of OX40-TC in lipid nanodiscs","authors":"Hao Wang , Xiaohui Zhang , Cunlong Yin , Lu Liu , Liang Chen , Hengfang Tang , Bo Wu , Junfeng Wang","doi":"10.1016/j.pep.2025.106697","DOIUrl":"10.1016/j.pep.2025.106697","url":null,"abstract":"<div><div>Tumor necrosis factor receptor superfamily member 4 (OX40) is essential for the activation and maintenance of T cell immune function because of its recognition and regulation in signal transduction, therefore understanding the structure and dynamics of the membrane protein OX40 in a near-native membrane environment is critical for elucidating its functions. However, efforts to prepare OX40 in a stable, functional form and reconstitute it into a membrane-mimicking lipid system face persistent challenges, limiting progress in its structural and functional characterization. Here, we developed an efficient method for the expression in <em>E. coli</em> and purification of the N-terminal truncated transmembrane and cytoplasmic domains of OX40 (OX40-TC) using a TrpLE fusion system. High-purity OX40-TC was achieved and successfully reconstituted into membrane scaffold protein (MSP)-nanodiscs for the first time. Characterization through size-exclusion chromatography, dynamic light scattering, and transmission electron microscopy confirmed their stability and homogeneity. This study provides a reliable platform for investigating the structure and function of OX40-TC, advancing our understanding of OX40-mediated signaling and its therapeutic potential.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106697"},"PeriodicalIF":1.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143551522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Mini review for niche downstream processes","authors":"Tsutomu Arakawa , Daisuke Ejima , Yui Tomioka , Chiaki Sakuma , Teruo Akuta","doi":"10.1016/j.pep.2025.106690","DOIUrl":"10.1016/j.pep.2025.106690","url":null,"abstract":"<div><div>We review here several niche downstream purification processes that are not covered by other articles in this special issue. The first is the use of activated carbon to capture contaminants and clarify culture medium for purification of proteins, including antibodies. Flow-through operation of the activated carbon filter showed over 80 % recovery of antibodies with 10--fold reduction of host cell proteins and effective lipopolysaccharide and virus removal. The second is salt or arginine-tolerant column chromatography, in which the respective resins, such as hydroxyapatite and mixed-mode resins, can bind proteins in the presence of salt or arginine at high concentrations. Effective use of arginine was also suggested in size exclusion and affinity chromatography. The last is the isolation of proteins using gel electrophoresis and simple extraction procedure. These technologies offer simple and cost-effective methods for purifying proteins and protein complexes in the native state.</div></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"230 ","pages":"Article 106690"},"PeriodicalIF":1.4,"publicationDate":"2025-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}