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Activation of three natural product biosynthetic gene clusters from Streptomyces lavendulae CGMCC 4.1386 by a reporter-guided strategy

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Synthetic and Systems Biotechnology 2018年第4期3卷 254-260页

作者： Pengwei Li Zhengyan Guo Wei Tang Yihua Chen State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing100101China University of Chinese Academy of Sciences Beijing100049China

Along with the fast developing of DNA sequencing technology,a great number of natural product biosynthetic gene clusters have been discovered by bioinformatic analysis,which demands novel high-throughput genome mining methods to obtain the diverse compounds dictated by those gene clusters.In this work,a method based on the reporter gene xylE was established to screen for the activation conditions of thirteen different gene clusters from Streptomyces lavendulae CGMCC 4.1386.In this reporter-guided method,the key structure gene was replaced by a xylE-kanaR cassette with the xylE gene being controlled by the transcription and translation machinery of the key structure gene.It not only facilitated the screening of activation conditions,but also provided the null mutants of specific natural product gene clusters as controls to link those clusters with their products conveniently.The potential activation conditions of eleven gene clusters from S.lavendulae CGMCC 4.1386 were obtained.In addition,activation of three of the eleven gene clusters was confirmed and their products were identified.

关键词： Natural product Genome mining Streptomyces reporter

Tetracysteine as a reporter for Gene Therapy

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Biomedical and Environmental Sciences 2009年第6期22卷 496-501页

作者： CHEN-YU XU YING GU WANG-HENG HOU YU-QIONG QUE SHUANG-GUAN GAO TONG CHENG NING-SHAO XI National Institute of Diagnostics and Vaccine Development for Infectious Diseases School of Life Sciences Xiamen University Xiamen 361005 Fujian China Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering School of Life Sciences Xiamen University Xiamen 361005 Fujian China

Objective To study the feasibility of using tetracysteine （TC） reporter in gene therapy. Methods Effects of TC reporter and conventional reporter genes encoding green fluorescence protein （GFP） and luciferase （Luc） on expression and function of the therapeutic gene MGMT^P140K were compared. Cytotoxicity and drug resistance were studied by Western blot. TC reporter used in therapy was analyzed by flow cytometry （FCM）. Results The TC reporter had no toxicity to cells and neither affected the expression or activity of therapeutic gene as compared to GFP and Luc. TC could be used in blood sample detection. Conclusion TC is a new kind of reporter gene for lentiviral vector in future gene therapy.

关键词： Tetracysteine Lentiviral vector reporter Gene therapy

A reporter for noninvasively monitoring gene expression and plant transformation

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Horticulture Research 2020年第1期7卷 821-826页

作者： Yubing He Tao Zhang Hui Sun Huadong Zhan Yunde Zhao State Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural UniversityNanjing 210095China National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research(Wuhan) Huazhong Agricultural UniversityWuhan 430070China Section of Cell and Developmental Biology University of California San Diego9500 Gilman DriveLa JollaCA 92093-0116USA Tata Institute for Genetics and Society-UCSD La JollaCA 92093-0335USA

reporters have been widely used to visualize gene expression,protein localization,and other cellular activities,but the commonly used reporters require special equipment,expensive chemicals,or invasive treatments.Here,we construct a new reporter RUBY that converts tyrosine to vividly red betalain,which is clearly visible to naked eyes without the need of using special equipment or chemical treatments.We show that RUBY can be used to noninvasively monitor gene expression in plants.Furthermore,we show that RUBY is an effective selection marker for transformation events in both rice and Arabidopsis.The new reporter will be especially useful for monitoring cellular activities in large crop plants such as a fruit tree under field conditions and for observing transformation and gene expression in tissue culture under sterile conditions.

关键词： transformation reporter sterile

Overview of the reporter genes and reporter mouse models

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Animal Models and Experimental Medicine 2018年第1期1卷 29-35页

作者： Shun Li Li-xiang Chen Xiu-hua Peng Chao Wang Bo-yin Qin Dan Tan Cheng-xiao Han Hua Yang Xiao-nan Ren Fang Liu Chun-hua Xu Xiao-hui Zhou Key Laboratory of Medical Molecular Virology Shanghai Public Health Clinical Center Ministry of Education and HealthFudan University School of Agriculture and Biology Shanghai Jiao Tong University

reporter genes are widely applied in biotechnology and biomedical research owning to their easy observation and lack of toxicity. Taking advantage of the reporter genes in conjunction with imaging technologies, a large number of reporter mouse models have been generated. reporter mouse models provide systems that enable the studies of live cell imaging, cell lineage tracing, immunological research and cancers etc. in vivo. In this review, we describe the types of different reporter genes and reporter mouse models including, random reporter strains, Cre reporter strains and ROSA26 reporter strains. Collectively, these reporter mouse models have broadened scientific inquires and provided potential strategies for generation of novel reporter animal models with enhanced capabilities.

关键词： Cre reporter strains random reporter strains reporter genes ROSA26 reporter strains

reporter Genes: A Practical Guide (Methods in Molecular Biology) 1

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Tppp3~+synovial/tendon sheath progenitor cells contribute to heterotopic bone after trauma

丛书名： Methods in Molecular Biology 411

2007年

作者： Siobhan Loughna and Deborah Henderson Philip A. Seymour and Maike Sander Maria Limberis Peter Bell and James M. Wilson Wolfgang J. Weninger and Timothy J. Mohun George H. Patterson Karin Edoff James S. Dods and Andrea H. Brand Charles G. Bailey and

ISBN: (纸本)9781588297396

reporter genes have played, and continue to play, a vital role in many areas of biological research by providing a ready means for qualitative and quantitative assessment of the activity of genes and location of gene products in different environments. This book describes practical protocols for experimentation with the most useful reporter genes for mammalian systems that are available

关键词： Genes reporter

来源：畅想之星图书光盘版（南... 评论

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骨研究:英文版 2023年第3期11卷 548-559页

作者： Ji-Hye Yea Mario Gomez-Salazar Sharon Onggo Zhao Li Neelima Thottappillil Masnsen Cherief Stefano Negri Xin Xing Qizhi Qin Robert Joel Tower Chen-Ming Fan Benjamin Levi Aaron W.James Department of Pathology Johns Hopkins UniversityBaltimoreMD 21205USA Orthopaedic and Trauma Surgery Unit Department of SurgeryDentistryPaediatrics and Gynaecology of the University of VeronaVeronaItaly Center for Organogenesis and Trauma Department of SurgeryUniversity of Texas SouthwesternDallasTXUSA Carnegie Institution for Science BaltimoreMDUSA

Heterotopic ossification(HO)is a pathological process resulting in aberrant bone formation and often involves synovial lined tissues.During this process,mesenchymal progenitor cells undergo endochondral ossification.Nonetheless,the specific cell phenotypes and mechanisms driving this process are not well understood,in part due to the high degree of heterogeneity of the progenitor cells involved.Here,using a combination of lineage tracing and single-cell RNA sequencing(sc RNA-seq),we investigated the extent to which synovial/tendon sheath progenitor cells contribute to heterotopic bone formation.For this purpose,Tppp3(tubulin polymerization-promoting protein family member 3)-inducible reporter mice were used in combination with either Scx(Scleraxis)or Pdgfra(platelet derived growth factor receptor alpha)reporter mice.Both tendon injury-and arthroplasty-induced mouse experimental HO models were utilized.Sc RNA-seq of tendon-associated traumatic HO suggested that Tppp3 is an early progenitor cell marker for either tendon or osteochondral cells.Upon HO induction,Tppp3 reporter^(+)cells expanded in number and partially contributed to cartilage and bone formation in either tendon-or joint-associated HO.In double reporter animals,both Pdgfra^(+)Tppp3^(+)and Pdgfra^(+)Tppp3^(-) progenitor cells gave rise to HO-associated cartilage.Finally,analysis of human samples showed a substantial population of TPPP3^(-) expressing cells overlapping with osteogenic markers in areas of heterotopic bone.Overall,these data demonstrate that synovial/tendon sheath progenitor cells undergo aberrant osteochondral differentiation and contribute to HO after trauma.

关键词： progenitor tendon reporter

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STYLE RADAR

城市漫步（GBA版） 2024年第1期 11-11页

"Thank you foryour support.Ilove you China,my good friend!"...excaimed Tennis World Champion Novak Djokovic,to everyone's astonish-ment,in excellent Chinese.At the United Cup in Perth,Novak Djokovic surprised a Chines... 详细信息

关键词： everyone reporter friend

mgr-mir-9 implicates Meloidogyne graminicola infection in rice by targeting the effector MgPDI

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Journal of Integrative Agriculture 2023年第5期22卷 1445-1454页

作者： TIAN Zhong-ling ZHOU Jia-yan ZHENG Jing-wu HAN Shao-jie Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province Interdisciplinary Research Academy(IRA)Zhejiang Shuren UniversityHangzhou 310015P.R.China Laboratory of Plant Nematology Institute of BiotechnologyCollege of Agriculture&BiotechnologyZhejiang UniversityHangzhou 310058P.R.China Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insect Pests Zhejiang UniversityHangzhou 310058P.R.China

MicroRNAs (miRNAs),a class of small non-coding RNAs,are crucial endogenous gene regulators in a range of animals,including plant-parasitic nematodes.Meloidogyne graminicola is an obligate sedentary endoparasite of rice and causes significant yield losses.A number of studies focused on the roles of M.graminicola effectors during the parasitic process;however,how nematode miRNAs regulate its effectors needs elucidating.In this research,we analyzed a cluster of M.graminicola miRNAs obtained at the second-stage juveniles (J2s) stage that are closely linked to the regulation of M.graminicola effectors.There are 49 767 105 total clean reads obtained from three libraries.A total of 233 known miRNAs and 21 novel miRNAs were identified.Among the known miRNAs,mgr-lin-4,mgr-mir-1,mgr-mir-100,mgrmir-86,mgr-mir-279,mgr-mir-87,mgr-mir-71,mgr-mir-9,mgr-mir-50,mgr-mir-72,and mgr-mir-34 are the most abundant11 miRNAs families.Moreover,the expression levels of selected miRNAs were validated by real-time quantitative PCR.We hypothesized that these miRNAs might regulate the expression of secreted effectors during the J2s stage to facilitate its infection.Consistent with this,we found that mgr-mir-9 targets MgPDI,an important M.graminicola effector mRNA.In addition to that,J2s treated with mgr-mir-9 mimics showed down-regulation of MgPDI expression and reduced reproductive ability,alluding mgr-mir-9 is involved in nematode infection.These results provide novel insight into the regulatory functions of M.graminicola miRNAs during the infection and identify miRNAs and their effector targets as potential key management targets to limit parasite survival during the early stages of infection.

关键词： microRNA function Meloidogyne graminicola deep sequencing Dual-Luciferase reporter Assay System proteindisulfideisomerase

Alpha-fetoprotein-targeted reporter gene expression imaging in hepatocellular carcinoma

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World Journal of Gastroenterology 2016年第27期22卷 6127-6134页

作者： Kwang Il Kim Hye Kyung Chung Ju Hui Park Yong Jin Lee Joo Hyun Kang Molecular Imaging Research Center Korea Institute of Radiological and Medical Sciences Korea Drug Development Platform using Radio-Isotope Korea Institute of Radiological and Medical Sciences

Hepatocellular carcinoma(HCC) is one of the most common cancers in Eastern Asia, and its incidence is increasing globally. Numerous experimental models have been developed to better our understanding of the pathogenic mechanism of HCC and to evaluate novel therapeutic approaches. Molecular imaging is a convenient and up-to-date biomedical tool that enables the visualization, characterization and quantification of biologic processes in a living subject. Molecular imaging based on reporter gene expression, in particular, can elucidate tumor-specific events or processes by acquiring images of a reporter gene's expression driven by tumor-specific enhancers/promoters. In this review, we discuss the advantages and disadvantages of various experimental HCC mouse models and we present in vivo images of tumorspecific reporter gene expression driven by an alphafetoprotein(AFP) enhancer/promoter system in a mouse model of HCC. The current mouse models of HCC development are established by xenograft, carcinogen induction and genetic engineering, representing the spectrum of tumor-inducing factors and tumor locations. The imaging analysis approach of reporter genes driven by AFP enhancer/promoter is presented for these different HCC mouse models. Such molecular imaging can provide longitudinal information about carcinogenesis and tumor progression. We expect that clinical application of AFP-targeted reporter gene expression imaging systems will be useful for the detection of AFP-expressing HCC tumors and screening of increased/decreased AFP levels due to disease or drug treatment.

关键词： Alpha-fetoprotein Hepatocellular carcinoma Molecular imaging reporter gene Tumor-specific enhancer/p