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Repressor elements provide insights into tissue development and phenotypes in pigs

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Zoological Research 2024年第6期45卷 1313-1326页

作者： Yue-Dong Zhang Chao Guo Hang Liu Yun Gao yongjun tan Longjian Niu Ligang Wang Lixian Wang David MIrwin Chunhui Hou Zhong-Yin Zhou Ya-Ping Zhang Key Laboratory of Genetic Evolution&Animal Models and Yunnan Key Laboratory of Molecular Biology of Domestic Animals Kunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650223China Kunming College of Life Science University of Chinese Academy of SciencesKunmingYunnan 650204China State Key Laboratory for Conservation and Utilization of Bio-resources in Yunnan Yunnan UniversityKunmingYunnan 650091China School of Life Science Yunnan UniversityKunmingYunnan 650091China School of Life Sciences Division of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhui 230026China State Key Laboratory of Hybrid Rice Hunan Hybrid Rice Research CenterHunan Academy of Agricultural SciencesChangshaHunan 410125China School of Public Health and Emergency Management Southern University of Science and TechnologyShenzhenGuangdong 518055China Shenzhen Key Laboratory of Cardiovascular Health and Precision Medicine Southern University of Science and TechnologyShenzhenGuangdong 518055China Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation of Ministry of Agriculture Institute of Animal ScienceChinese Academy of Agricultural SciencesBeijing 100081China Department of Laboratory Medicine and Pathobiology University of TorontoToronto M5S 1A8Canada Yunnan Key Laboratory of Biodiversity Information KIZ-CUHK Joint Laboratory of Bioresources and Molecular Research in Common Diseases Kunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650223China Center for Excellence in Animal Evolution and Genetics Chinese Academy of SciencesKunmingYunnan 650223China

Repressor elements significantly influence economically relevant phenotypes in pigs;however,their precise roles and characteristics are inadequately *** the present study,we employed H3K27me3 profiling,assay for transposase-accessible chromatin with highthroughput sequencing(ATAC-seq),and RNA sequencing(RNA-seq)data across six tissues derived from three embryonic layers to identify and map 2034 super repressor elements(SREs) and 22223 typical repressor elements(TREs) in the pig ***,many repressor elements were conserved across mesodermal and ectodermal *** exhibited tight regulation of their target genes,affecting a limited number of genes within a specific genomic region with pronounced effects,while TREs exerted broader but weaker regulation over a wider range of target ***,in neuronal tissues,genes regulated by repressor elements started to be repressed during the differentiation of stem cells into progenitor ***,analysis showed that many repressor elements exhibited cooperative and additive effects on the modulation of KLF4 *** research provides the first comprehensive map of pig repressor elements,serving as an essential reference for future studies on repressor elements.

关键词： Repressor elements Super repressor elements Silencer Regulatory model Tissue development KLF4 Pig

Bst DNA聚合酶的纯化及等温扩增检测体系的构建

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湖南大学学报（自然科学版） 2019年第12期46卷 98-106页

作者：谭拥军吴莎莎谭桂湘向勤湖南大学生物学院

Bst DNA聚合酶是等温扩增反应中的一个关键酶,但由于专利限制,目前国内实验室所使用的Bst DNA聚合酶大多需要从国外进口,价格昂贵且长途运输影响酶活性,因此亟需自主生产出Bst DNA聚合酶应用于等温扩增检测技术.通过构建Bst Fragement... 详细信息

Bst DNA聚合酶是等温扩增反应中的一个关键酶,但由于专利限制,目前国内实验室所使用的Bst DNA聚合酶大多需要从国外进口,价格昂贵且长途运输影响酶活性,因此亟需自主生产出Bst DNA聚合酶应用于等温扩增检测技术.通过构建Bst Fragement的原核表达质粒,采用原核蛋白表达系统和His-Tag亲和纯化手段,制备成本低,活性高,特异性强的Bst DNA聚合酶.同时,针对特定的模板,对自制Bst DNA聚合酶的扩增反应进行了进一步优化,用不同的产物鉴定方法对扩增效果进行检测,并最终筛选出该酶最优的扩增反应条件为60 mM K^+、30 mM(NH4)2SO4、pH为9.0.最后,验证了Bst DNA聚合酶能用于快速准确检测肺炎支原体、肺炎衣原体,降低了相关病原体核酸检测的成本,为之后核酸检测在基层医疗的更广泛应用提供了条件.

关键词： Bst DNA聚合酶重组蛋白扩增核酸检测

The metal tolerance protein OsMTP11 facilitates cadmium sequestration in the vacuoles of leaf vascular cells for restricting its translocation into rice grains

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Molecular Plant 2024年第11期17卷 1733-1752页

作者： Peng Liu Liang Sun Yu Zhang yongjun tan Yuxing Zhu Can Peng Jiurong Wang Huili Yan Donghai Mao Guohua Liang Gang Liang Xiaoxiang Li Yuntao Liang Feng Wang Zhenyan He Wenbang tang Daoyou Huang Caiyan Chen Key Laboratory of Agro-Ecological Processes in Subtropical Region Institute of Subtropical AgricultureChinese Academy of SciencesChangsha 410125China College of Agronomy Hunan Agricultural UniversityChangsha 410128China Key Laboratory of Plant Resources Institute of BotanyChinese Academy of SciencesBeijing 100093China Jiangsu Key Laboratory of Crop Genetics and Physiology Yangzhou UniversityYangzhou 225009China CAS Key Laboratory of Tropical Plant Resources and Sustainable Use Xishuangbanna Tropical Botanical GardenChinese Academy of SciencesKunmingYunnan 650223China Hunan Rice Research Institute Hunan Academy of Agricultural ScienceChangsha 410125China Guangxi Key Laboratory of Rice Genetics and Breeding Rice Research InstituteGuangxi Academy ofAgricultural SciencesNanning 530007China University of Chinese Academy of Sciences Beijing 100049China

Rice(Oryza sativa)provides>20%of the consumed calories in the human ***,rice is also a leading source of dietary cadmium(Cd)that seriously threatens human *** the genetic network that underlies the grain-Cd accumulation will benefit the development of low-Cd rice and mitigate the effects of Cd accumulation in the rice *** this study,we identified a QTL gene,OsCS1,which is allelic to OsMTP11 and encodes a protein sequestering Cd in the leaf during vegetative growth and preventing Cd from being translocated to the grain after heading in ***1 is predominantly expressed in leaf vascular parenchyma cells,where it binds to a vacuole-sorting receptor protein OsVSR2 and is translocated intracellularly from the trans-Golgi network to pre-vacuolar compartments and then to the *** this trafficking process,OsCS1 actively transports Cd into the endomembrane system and sequesters it in the *** are natural variations in the promoter of OsCS1 between the indica and japonica rice *** of a G-box-like motif in the promoter region of the superior allele of OsCS1 from indica rice enhances the binding of the transcription factor OsIRO2 to the OsCS1 promoter,thereby promoting OsCS1 *** of this allele into commercial rice varieties could significantly lower grain-Cd levels compared to the inferior allele present in japonica ***,our findings offer new insights into the genetic control of leaf-to-grain Cd translocation and provide a novel gene and its superior allele for the genetic improvement of low-Cd variety in rice.

关键词： rice OsCS1/OsMTP11 grain Cd TGN–PVC–vacuole transport allelic variation

R9-FOXM1(1-234aa)重组蛋白批量纯化及其抑瘤效应研究

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湖南大学学报（自然科学版） 2017年第12期44卷 116-120页

作者：谭拥军余景卫陈燕向勤谭桂湘湖南大学生物学院湖南长沙410082

通过基因工程构建重组表达质粒pET15b-R9-FOXM1(1-234aa),转化大肠杆菌建立构建表达R9-FOXM1(1-234aa)的菌株.采用原核表达系统和His-tag亲和纯化手段,规模化制备纯化穿膜肽R9-FOXM1(1-234aa),获得的蛋白的纯度达到90%以上.用R9-FOXM1(1-234aa)穿膜肽处理不同的肿瘤细胞,通过MTT实验研究其细胞效应.结果显示:当R9-FOXM1(1-234aa)穿膜肽的浓度达到2mM时,肿瘤细胞的死亡率为50%左右.实验表明穿膜肽R9-FOXM1(1-234aa)抑制不同肿瘤细胞的生长,有可能成为治疗肿瘤的潜在蛋白类药物.

关键词：多聚精氨酸细胞穿膜肽 FOXM1 肿瘤治疗

E-cadherin启动子的荧光示踪体系的建立及验证

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湖南大学学报（自然科学版） 2020年第12期47卷 131-136页

作者：谭拥军吴东丽余雳陈燕湖南大学生物学院湖南长沙410082

为实现对肺癌A549细胞的EMT示踪,构建了E-cadherin基因启动子的慢病毒质粒,并与包装质粒pVSVG、Δ8.91共转染HEK293T细胞,包装成有活性的慢病毒,收取病毒感染A549细胞,利用流式细胞仪和Western blot检测感染成功.进一步用TGF-β诱导感... 详细信息

为实现对肺癌A549细胞的EMT示踪,构建了E-cadherin基因启动子的慢病毒质粒,并与包装质粒pVSVG、Δ8.91共转染HEK293T细胞,包装成有活性的慢病毒,收取病毒感染A549细胞,利用流式细胞仪和Western blot检测感染成功.进一步用TGF-β诱导感染成功的A549细胞,通过在荧光显微镜下观察荧光变化、荧光定量PCR和Western blot检测GFP和E-cadherin的变化,发现在TGF-β诱导48 h后,A549细胞形态由鹅卵石样变成长梭形.并且荧光显微镜观察到相比较于未诱导的细胞,诱导后的A549细胞绿色荧光明显变暗,同时GFP和E-cadherin的RNA水平和蛋白质表达水平均降低.以E-cadherin基因启动子为基础建立了A549细胞的EMT荧光示踪体系,为进一步研究肺癌的EMT过程提供了材料.

关键词：启动子 E-cadherin 示踪肺癌细胞

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细胞选择性穿膜肽的穿膜效果研究

湖南大学学报（自然科学版） 2019年第6期46卷 83-87页

作者：谭拥军王坤余雳黄小芹陈燕谭桂湘湖南大学生物学院

细胞穿膜肽是一类能够穿透细胞膜并且不损坏细胞膜结构的小分子多肽,该多肽一般由不多于30个氨基酸组成.针对FITC标记的具有肺癌或肝癌细胞选择性的两种穿膜肽,设置不同浓度梯度,分别处理多种肿瘤细胞,荧光显微镜下观察不同作用浓度下... 详细信息

细胞穿膜肽是一类能够穿透细胞膜并且不损坏细胞膜结构的小分子多肽,该多肽一般由不多于30个氨基酸组成.针对FITC标记的具有肺癌或肝癌细胞选择性的两种穿膜肽,设置不同浓度梯度,分别处理多种肿瘤细胞,荧光显微镜下观察不同作用浓度下细胞中绿色荧光的强弱,并通过流式细胞仪检测含有荧光的细胞数.结果表明:两种细胞穿膜肽具有不同的细胞选择性.CPP33在作用浓度为10μmol/L时能观察到较强的荧光选择性,流式细胞仪检测含有荧光的细胞数约大于50%,CPP44在作用浓度为10μmol/L时能观察到较强的荧光选择性,流式细胞仪检测含有荧光的细胞数约大于60%.并且随着作用浓度的增加,两种细胞中含有荧光的细胞数不断上升.因此这种针对肿瘤细胞的选择性穿膜肽可能为小分子药物在体内的精确递送提供一种新的方法.

关键词：肿瘤选择性穿膜肽肺癌肝癌浓度梯度小分子药物

慢病毒介导MCF-7细胞中FOXP2基因沉默体系的建立

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湖南大学学报（自然科学版） 2018年第6期45卷 128-132页

作者：谭拥军郭明月向勤李谕湖南大学生物学院湖南长沙410082

为了实现FOXP2基因在MCF-7细胞中稳定低表达,采用基因工程的方法构建pMAGic7.1-shFOXP2慢病毒干扰载体,并将其与辅助质粒共转染HEK293T细胞,包装shFOXP2干扰慢病毒,收取病毒上清加入到MCF-7细胞中,培养48h,荧光定量PCR检测FOXP2的基因m... 详细信息

为了实现FOXP2基因在MCF-7细胞中稳定低表达,采用基因工程的方法构建pMAGic7.1-shFOXP2慢病毒干扰载体,并将其与辅助质粒共转染HEK293T细胞,包装shFOXP2干扰慢病毒,收取病毒上清加入到MCF-7细胞中,培养48h,荧光定量PCR检测FOXP2的基因mRNA表达水平,Western Blotting检测FOXP2蛋白表达水平.获得了pMAGic7.1-shFOXP2干扰载体,从而成功建立FOXP2基因稳定干扰的MCF-7细胞株,为进一步研究FOXP2在乳腺癌发生发展中的功能提供了研究材料.

关键词： FOXP2 慢病毒载体 RNA干扰乳腺癌MCF-7细胞

城市道路面层必须引起重视的几种通病分析及防治

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城市道桥与防洪 2010年第11期 79-81页

作者：谭永军镇江市市政设施管理处江苏镇江212001

该文介绍了城市道路面层施工工艺及施工过程中产生的质量通病的防治、控制措施。同时针对施工后发现的病害也提出了相应处治措施。

关键词：道路面层病害防治措施

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抗肿瘤多肽M1-21的剂型优化

湖南大学学报（自然科学版） 2022年第6期49卷 146-151页

作者：谭拥军裴超柱程浩杰卜会铜谭桂湘邓磊湖南大学生物学院湖南长沙410082

通过去溶剂化方法,将单体抗肿瘤多肽M1-21纳米化,形成50 nm左右大小的多肽纳米颗粒(Nano-M1-21).运用动态光色散技术(DLS)和低压透射电镜(TEM)对Nano-M1-21进行表征,并进行体外37℃条件下颗粒稳定性测试.运用乳腺癌MDA-MB-231、MCF-7和... 详细信息

通过去溶剂化方法,将单体抗肿瘤多肽M1-21纳米化,形成50 nm左右大小的多肽纳米颗粒(Nano-M1-21).运用动态光色散技术(DLS)和低压透射电镜(TEM)对Nano-M1-21进行表征,并进行体外37℃条件下颗粒稳定性测试.运用乳腺癌MDA-MB-231、MCF-7和4T1细胞验证Nano-M1-21的抗肿瘤效果,结果表明Nano-M1-21更容易被细胞摄取;相比单体多肽,达到抑制肿瘤细胞的浓度更低.运用小鼠乳腺癌4T1细胞活体肿瘤移植模型,发现Nano-M1-21展示出良好的抑瘤效果.总之,单体肽M1-21经纳米剂型优化后,有利于改善细胞的摄取,显示出更好的抑瘤效果.

关键词： M1-21 Nano-M1-21 纳米化抑瘤效果多肽