Synthetic and Systems Biotechnology最新文献

{"title":"Design and engineering of logic genetic-enzymatic gates based on the activity of the human CYP2C9 enzyme in permeabilized Saccharomyces cerevisiae cells","authors":"Rana Azeem Ashraf,&nbsp;Matthias Bureik,&nbsp;Mario Andrea Marchisio","doi":"10.1016/j.synbio.2024.03.013","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.013","url":null,"abstract":"<div><p>Gene circuits allow cells to carry out complex functions such as the precise regulation of biological metabolic processes. In this study, we combined, in the yeast <em>S. cerevisiae</em>, genetic regulatory elements with the enzymatic reactions of the human CYP2C9 and its redox partner CPR on luciferin substrates and diclofenac. <em>S. cerevisiae</em> cells were permeabilized and used as enzyme bags in order to host these metabolic reactions. We engineered three different (genetic)-enzymatic basic Boolean gates (YES, NOT, and N-IMPLY). In the YES and N-IMPLY gates, human CYP2C9 was expressed under the galactose-inducible <em>GAL1</em> promoter. The carbon monoxide releasing molecule CORM-401 was used as an input in the NOT and N-IMPLY gates to impair CYP2C9 activity through inhibition of the Fe⁺²- heme prosthetic group in the active site of the human enzyme. Our study provides a new approach in designing synthetic bio-circuits and optimizing experimental conditions to favor the heterologous expression of human drug metabolic enzymes over their endogenous counterparts. This new approach will help study precise metabolic attributes of human P450s.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000474/pdfft?md5=1b0e86900ca2e71dc42b1d5d44a1c2c4&pid=1-s2.0-S2405805X24000474-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140332469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Promoting the production of challenging proteins via induced expression in CHO cells and modified cell-free lysates harboring T7 RNA polymerase and mutant eIF2α","authors":"Jeffrey L. Schloßhauer ,&nbsp;Lena Tholen ,&nbsp;Alexander Körner ,&nbsp;Stefan Kubick ,&nbsp;Sofia Chatzopoulou ,&nbsp;Anja Hönow ,&nbsp;Anne Zemella","doi":"10.1016/j.synbio.2024.03.011","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.011","url":null,"abstract":"<div><p>Chinese hamster ovary (CHO) cells are crucial in biopharmaceutical production due to their scalability and capacity for human-like post-translational modifications. However, toxic proteins and membrane proteins are often difficult-to-express in living cells. Alternatively, cell-free protein synthesis can be employed. This study explores innovative strategies for enhancing the production of challenging proteins through the modification of CHO cells by investigating both, cell-based and cell-free approaches. A major result in our study involves the integration of a mutant eIF2 translation initiation factor and T7 RNA polymerase into CHO cell lysates for cell-free protein synthesis. This resulted in elevated yields, while eliminating the necessity for exogenous additions during cell-free production, thereby substantially enhancing efficiency. Additionally, we explore the potential of the <em>Rosa26</em> genomic site for the integration of T7 RNA polymerase and cell-based tetracycline-controlled protein expression. These findings provide promising advancements in bioproduction technologies, offering flexibility to switch between cell-free and cell-based protein production as needed.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000450/pdfft?md5=b72c35cbaf02c78cd3e50f72c48774d1&pid=1-s2.0-S2405805X24000450-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140345128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined strategies for improving the heterologous expression of a novel xylanase from Fusarium oxysporum Fo47 in Pichia pastoris","authors":"Chun Liu ,&nbsp;Yaping Zhang ,&nbsp;Chunting Ye ,&nbsp;Fengguang Zhao ,&nbsp;Yian Chen ,&nbsp;Shuangyan Han","doi":"10.1016/j.synbio.2024.03.012","DOIUrl":"10.1016/j.synbio.2024.03.012","url":null,"abstract":"<div><p>Xylanase, an enzyme capable of hydrolyzing non-starch polysaccharides found in grain structures like wheat, has been found to improve the organizational structure of dough and thus increase its volume. In our past work, one promising xylanase FXYL derived from <em>Fusarium oxysporum</em> Fo47 and first expressed 779.64 U/mL activity in <em>P. pastoris</em>. It has shown significant potential in improving the quality of whole wheat bread, making it become a candidate for development as a new flour improver. After optimization of expression elements and gene dose, the xylanase activity of FXYL strain carrying three-copies reached 4240.92 U/mL in <em>P. pastoris</em>. In addition, 12 factors associated with the three stages of protein expression pathway were co-expressed individually in order in three-copies strain, and the translation factor Pab1 co-expression increased FXYL activity to 8893.53 U/mL. Nevertheless, combining the most effective or synergistic factors from three stages did not exhibit better results than co-expressing them alone. To further evaluate the industrial potential, the xylanase activity and protein concentration reached 81184.51 U/mL and 11.8 g/L in a 5 L fed-batch fermenter. These engineering strategies improved the expression of xylanase FXYL by more than 104-fold, providing valuable insights for the cost-effective industrial application of FXYL in the baking field.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000462/pdfft?md5=a333edf2d892452e44d608e5aca82730&pid=1-s2.0-S2405805X24000462-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140407083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Environment and synthetic biology","authors":"Hongzhi Tang,&nbsp;Maoyong Song","doi":"10.1016/j.synbio.2024.03.016","DOIUrl":"10.1016/j.synbio.2024.03.016","url":null,"abstract":"","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000504/pdfft?md5=783aeca9e59946a47fc8ee5ba7fac437&pid=1-s2.0-S2405805X24000504-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140274622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovering peptides and computational investigations of a multiepitope vaccine target Mycobacterium tuberculosis","authors":"Truc Ly Nguyen ,&nbsp;Heebal Kim","doi":"10.1016/j.synbio.2024.03.010","DOIUrl":"10.1016/j.synbio.2024.03.010","url":null,"abstract":"<div><p><em>Mycobacterium tuberculosis</em> (MTB) is the causative agent of tuberculosis (TB), a prevalent airborne infectious disease. Despite the availability of the Bacille Calmette-Guerin vaccine, its global efficacy remains modest, and tuberculosis persists as a significant global public health threat. Addressing this challenge and advancing towards the End MTB Strategy, we developed a multiepitope vaccine (MEV) based on immunoinformatics and computational approaches. Immunoinformatics screening of MBT protein identified immune-dominant epitopes based on Major Histocompatibility Complex (MHC) allele binding, immunogenicity, antigenicity, allergenicity, toxicity, and cytokine inducibility. Selected epitopes were integrated into an MEV construct with adjuvant and linkers, forming a fully immunogenic vaccine candidate. Comprehensive analyses encompassed the evaluation of immunological and physicochemical properties, determination of tertiary structure, molecular docking with Toll-Like Receptors (TLR), molecular dynamics (MD) simulations for all atoms, and immune simulations. Our MEV comprises 534 amino acids, featuring 6 cytotoxic T lymphocyte, 8 helper T lymphocyte, and 7 linear B lymphocyte epitopes, demonstrating high antigenicity and stability. Notably, molecular docking studies and triplicate MD simulations revealed enhanced interactions and stability of MEV with the TLR4 complex compared to TLR2. In addition, the immune simulation indicated the capacity to effectively induce elevated levels of antibodies and cytokines, emphasizing the vaccine's robust immunogenic response. This study presents a promising MEV against TB, exhibiting favorable immunological and physicochemical attributes. The findings provide theoretical support for TB vaccine development. Our study aligns with the global initiative of the End MTB Strategy, emphasizing its potential impact on addressing persistent challenges in TB control.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000449/pdfft?md5=b9a4b77cd51a2b1f0341ec4570d1e876&pid=1-s2.0-S2405805X24000449-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140272341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genus-specific secondary metabolome in Allokutzneria and Kibdelosporangium","authors":"Man-Jing Ma ,&nbsp;Wen-Chao Yu ,&nbsp;Huai-Ying Sun ,&nbsp;Bing-Cheng Dong ,&nbsp;Gang-Ao Hu ,&nbsp;Zhen-Yi Zhou ,&nbsp;Yi Hua ,&nbsp;Buddha Bahadur Basnet ,&nbsp;Yan-Lei Yu ,&nbsp;Hong Wang ,&nbsp;Bin Wei","doi":"10.1016/j.synbio.2024.03.015","DOIUrl":"10.1016/j.synbio.2024.03.015","url":null,"abstract":"<div><p>Rare actinomycete genera are highly recognized as a promising source of structurally diverse and bioactive natural products. Among these genera, <em>Allokutzneria</em> and <em>Kibdelosporangium</em> are two phylogenetically closely related and have been reported to encode some valuable biosynthetic enzymes and secondary metabolites. However, there is currently no relevant systematic research available to outline the linkage of genomic and metabolomics for specific secondary metabolites in these two promising genera. In this study, we first investigated the genus-specific secondary metabolic potential in <em>Allokutzneria</em> and <em>Kibdelosporangium</em> by comparing the diversity and novelty of their secondary metabolite biosynthetic gene clusters (BGCs). The specific secondary metabolites produced by two representative strains of these genera were comprehensively investigated using untargeted metabolomics techniques. The findings unveiled that the majority (95.4%) of the gene cluster families (GCFs) encoded by <em>Allokutzneria</em> and <em>Kibdelosporangium</em> were genus-specific, including NRPS GCFs encoding siderophores. The untargeted metabolomics analysis revealed that the metabolic profiles of two representative strains exhibit extensive specificity, with the culture medium having a big impact on the metabolic profiles. Besides, an MS-cluster featuring a series of hydroxamate-type siderophores was identified from <em>Allokutzneria albata</em> JCM 9917, with two of them, including a novel one (N-deoxy arthrobactin A), being experimentally validated. The present study offers valuable insights for the targeted discovery of genus-specific natural products from microorganisms.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000498/pdfft?md5=ee66342146f631def32737a4f8bca691&pid=1-s2.0-S2405805X24000498-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140274637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expanding catalytic promiscuity of a bifunctional terpene synthase through a single mutation-induced change in hydrogen-bond network within the catalytic pocket","authors":"Xinye Wang ,&nbsp;Yiyi Huang ,&nbsp;Weiyan Zhang ,&nbsp;Kangjie Lv ,&nbsp;Xiaoying Li ,&nbsp;Zhixin Wang ,&nbsp;Li Zhang ,&nbsp;Tom Hsiang ,&nbsp;Lixin Zhang ,&nbsp;Liming Ouyang ,&nbsp;Xueting Liu","doi":"10.1016/j.synbio.2024.03.007","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.007","url":null,"abstract":"<div><p>Fungal bifunctional terpene synthases (BFTSs) catalyze the formation of numerous di-/sester-/tri-terpenes skeletons. However, the mechanism in controlling the cyclization pattern of terpene scaffolds is rarely deciphered for further application of tuning the catalytic promiscuity of terpene synthases for expanding the chemical space. In this study, we expanded the catalytic promiscuity of <em>Fusarium oxysporum</em> fusoxypene synthase (FoFS) by a single mutation at L89, leading to the production of three new sesterterpenes. Further computational analysis revealed that the reconstitution of the hydrogen-bond (H-bond) network of second-shell residues around the active site of FoFS influences the orientation of the aromatic residue W69 within the first-shell catalytic pocket. Thus, the dynamic orientation of W69 alters the carbocation transport, leading to the production of diverse ring system skeletons. These findings enhance our knowledge on understanding the molecular mechanisms, which could be applied on protein engineering terpene synthases on regulating the terpene skeletons.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000413/pdfft?md5=613f6a2965129220b4e83fbec6870f86&pid=1-s2.0-S2405805X24000413-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140191858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering and finetuning expression of SerC for balanced metabolic flux in vitamin B6 production","authors":"Kai Chen ,&nbsp;Linxia Liu ,&nbsp;Jinlong Li ,&nbsp;Zhizhong Tian ,&nbsp;Hongxing Jin ,&nbsp;Dawei Zhang","doi":"10.1016/j.synbio.2024.03.005","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.005","url":null,"abstract":"<div><p>Vitamin B₆ plays a crucial role in cellular metabolism and stress response, making it an essential component for growth in all known organisms. However, achieving efficient biosynthesis of vitamin B₆ faces the challenge of maintaining a balanced distribution of metabolic flux between growth and production. In this study, our focus is on addressing this challenge through the engineering of phosphoserine aminotransferase (SerC) to resolve its redundancy and promiscuity. The enzyme SerC was semi-designed and screened based on sequences and predicted <em>k</em>_cat values, respectively. Mutants and heterologous proteins showing potential were then fine-tuned to optimize the production of vitamin B₆. The resulting strain enhances the production of vitamin B₆, indicating that different fluxes are distributed to the biosynthesis pathway of serine and vitamin B₆. This study presents a promising strategy to address the challenge posed by multifunctional enzymes, with significant implications for enhancing biochemical production through engineering processes.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000395/pdfft?md5=2bc29b99c91c574c90b1c87ff8cad95d&pid=1-s2.0-S2405805X24000395-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140290795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of an industrial bacterium Zymomonas mobilis for anaerobic l-serine production","authors":"Zhen Wang ,&nbsp;Xia Wang ,&nbsp;Xiongying Yan,&nbsp;Haixia Yi,&nbsp;Shuche He,&nbsp;Haoyu Zhang,&nbsp;Xinli Zhou,&nbsp;Qiaoning He,&nbsp;Shihui Yang","doi":"10.1016/j.synbio.2024.03.008","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.008","url":null,"abstract":"<div><p>Due to the complicated metabolic and regulatory networks of <span>l</span>-serine biosynthesis and degradation, microbial cell factories for <span>l</span>-serine production using non-model microorganisms have not been reported. In this study, a combination of synthetic biology and process optimization were applied in an ethanologenic bacterium <em>Zymomonas mobilis</em> for <span>l</span>-serine production. By blocking the degradation pathway while introducing an exporter EceamA from <em>E. coli</em>, <span>l</span>-serine titer in recombinant <em>Z. mobilis</em> was increased from 15.30 mg/L to 62.67 mg/L. It was further increased to 260.33 mg/L after enhancing the <span>l</span>-serine biosynthesis pathway. Then, 536.70 mg/L <span>l</span>-serine was achieved by removing feedback inhibition with a SerA mutant, and an elevated titer of 687.67 mg/L was further obtained through increasing s<em>erB</em> copies while enhancing the precursors. Finally, 855.66 mg/L <span>l</span>-serine can be accumulated with the supplementation of the glutamate precursor. This work thus not only constructed an <span>l</span>-serine producer to help understand the bottlenecks limiting <span>l</span>-serine production in <em>Z. mobilis</em> for further improvement, but also provides guidance on engineering non-model microbes to produce biochemicals with complicated pathways such as amino acids or terpenoids.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000425/pdfft?md5=a0a3c67c1dfcea572c0d2066d66a72a0&pid=1-s2.0-S2405805X24000425-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140180788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibacterial activity and mechanism of X33 antimicrobial oligopeptide against Acinetobacter baumannii","authors":"Qunlin Lu ,&nbsp;Xiaoyu Wu ,&nbsp;Yuan Fang ,&nbsp;Yuanxiu Wang ,&nbsp;Bin Zhang","doi":"10.1016/j.synbio.2024.03.002","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.03.002","url":null,"abstract":"<div><p><em>Acinetobacter baumannii</em> is a pathogenic bacterium widespread in human environments, especially in intensive care units, and is associated with high morbidity and infection rates. Multiple drug resistance in <em>A. baumannii</em> frequently leads to the death of patients, making the development of multi-effect antibacterial agents against this bacterium a research hotspot. We have previously found that the X33 antimicrobial oligopeptide can effectively inhibit the growth of <em>Penicillium digitatum</em> and <em>Candida albicans</em>. Herein, we evaluated the antibacterial activity of X33 antimicrobial oligopeptide against <em>A. baumannii</em> by determining the minimum inhibitory concentration, inhibition zone, and growth curve. The increase in extracellular alkaline phosphatase and the leakage of intracellular compounds confirmed the effect of X33 antimicrobial oligopeptide on the cell wall and membrane. Changes in reactive oxygen species, malondialdehyde, ATP, reducing sugar, soluble protein, and pyruvate content demonstrated that the incubation with X33 antimicrobial oligopeptide affected energy metabolism and oxidative stress. Consistent with the physiological characteristics, transcriptomics analysis indicated that incubation with X33 antimicrobial oligopeptide significantly induced changes in the expression of 2339 genes, including 1262 upregulated and 1077 downregulated genes, which participate in oxidative phosphorylation, ribosome, quorum sensing, fatty acid degradation, glycolysis/gluconeogenesis, and citrate cycle pathways. These results provide a fundamental basis for investigating the mechanism of X33 antimicrobial oligopeptide as a potential drug against <em>A. baumannii</em>.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X2400036X/pdfft?md5=8932767e05f3b3514a4d7afb84f9c31a&pid=1-s2.0-S2405805X2400036X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140160888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}