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Production of transgenic blastocyst by nuclear transfer from different types of somatic cells in cattle

Science china({1.fe Sciences) 2004年第2期47卷 183-189页

作者： GONG Guochun1, DAI Yunping1, FAN Baoliang1, ZHU Huabing2. WANG Haiping3, WANG lili1, FANG Changge1, WAN Rong3, liU Ying3, li Rong3 & li ning1 1. state key {2. for agrobiotechnology, china agricultural university, Beijing 100094,{12. 2. Institute of Animal Science, Chinese Academy of agricultural Sciences, Beijing 100094,{12. 3. Gentitan Biotechnology Ltd., Beijing 100084,{12. 1. state {1. {2. for {2. China Agricultural University 100094 Beijing China 2. Institute of Animal Science Chinese Academy of Agricultural Sciences 100094 Beijing China 3. Gentitan Biotechnology Ltd. 100084 Beijing China

The present study examined the effects of genetic manipulation to the donor cell and different types of transgenic donor cells on developmental potential of bovine nuclear trans-fer (NT) embryos. Four types of bovine somatic cells, including granulosa cells, fetal fibroblasts, fetal oviduct epithe{1.al cells and fetal ovary epithe{1.al cells, were transfected with a plasmid (pCE-EGFP-Ires-Neo-dNdB) contai{2. the enhanced green fluorescent protein (EGFP) and neomycin-resistant (Neor) genes by electroporation. After 1. days selection with 800 mg/mL G41., transgenic cell {1.nes from each type of somatic cells were obtained. Nontransgenic granulosa cells and all 4 types of transgenic somatic cells were used as nuclear donor to produce transgenic embryos by NT. There was no significant difference in development rates to the blas-tocyst stage for NT embryos from transgenic and nontransgenic granulosa cells (44.6% and 42.8%, respectively), and transfer of NT embryos derived from transgenic and nontransgenic granulosa cells to recipients resulted in similar pregnancy rates on day 90 (1.% and 2.%, re-spectively). The development rates to the blastocyst stage of NT embryos were significantly dif-ferent among different types of transgenic donor cells (Pduct epithe{1.al cell and granulosa cell (49.1. and 44.6%, respectively) were higher than those from fetal fibroblast (32.7%) and fetal ovary epithe{1.al cell (2..5%). These results suggest that (i) genetic manipulation to donor cells has no negative effect on in vitro and early in vivo develop-mental competence of bovine NT embryos and (ii) granulosa and fetal oviduct epithe{1.al cells can be used to produce transgenic bovine NT embryos more efficiently. In addition, GFP can be used to select transgenic NT embryos as a non-invasive selective marker.

关键词： transgenic, nuclear transfer, somatic cells, EGFP, blastocyst, bovine.

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单卵RT-PCR方法的优化和对牛卵母细胞中发育相关基因的表达研究

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生物化学与生物物理进展 2004年第7期31卷 659-665页

作者：李彦欣李世杰赵定胜赵春江杜卫华戴蕴平李宁中国农业大学农业生物技术国家重点实验室北京100094

到目前为止 ,虽然许多动物如羊、牛、猪、鼠、兔、马、骡等均已克隆成功 ,但克隆技术普遍存在着克隆效率低、流产率高、畸胎、巨胎和死亡率高等问题 .为了较全面地探索其基因方面的原因 ,需要对大量的候选基因进行分析 .而利用传统的RT ... 详细信息

到目前为止 ,虽然许多动物如羊、牛、猪、鼠、兔、马、骡等均已克隆成功 ,但克隆技术普遍存在着克隆效率低、流产率高、畸胎、巨胎和死亡率高等问题 .为了较全面地探索其基因方面的原因 ,需要对大量的候选基因进行分析 .而利用传统的RT PCR方法从单个卵母细胞或胚胎中获得的RNA量不足以分析大量的基因 .为了解决这一问题 ,利用并优化了一套全新的扩增细胞中整体cDNA的方法 ,使微量材料进行大量的基因表达分析成为现实 .通过该方法研究了与发育相关的重要基因IL6、IFτ、CX4 3、PSMC3、oct4和DNMT1.牛卵母细胞中的表达 .IL6、IFτ和CX4 3的表达与前人报告的结果相同 ,而DNMT1.PSMC3在牛卵母细胞中表达情况此前未见报道 .

关键词：单卵RT-PCR方法发育相关基因表达

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The excitation energy transfer between β-Car and Chla molecules in PS Ⅱ core antenna complexes CP43 and CP47

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Chinese Science Bulletin 2000年第17期45卷 1579-1583页

作者： SHAN Jixiu, WANG Jushuo, li liangbi,ZHAO Nanming & KUANG Tingyun1. {2. of Photosynthesis Basic Research, Institute of Botany, Chinese Academy of Sciences, Beijing 1.0093, china 2. Department of Biological Sciences and Biotechnology, {2. key {2. of Biomembrane and Membrane Biotechnology, Tsinghua university, Beijing 1.0084, chinaCorrespondence should be addressed to Kuang Tingyun laboratory of Photosynthesis Basic Research Institute of Botany Chinese Academy of Sciences Beijing China Department of Biological Sciences and Biotechnology State Key Laboratory of Biomembrane and Membrane Biotechnology Tsinghua University Beijing China

The excitation energy transfer efficiency between p-Car and Chla molecules in purified CP43 and CP47 was calculated by comparing absorption and fluorescence excitation after normalization at 550 nm, CP43 had an energy transfer efficiency of {1..1. while the CP47 had an energy transfer efficiency of 6{1.8%, proving that excitation energy was transferred between p-Car and Chla molecules in CP43 and CP47 at normal conditions. The excitation energy transfer between p-Car and Chla molecules in CP43 and CP47 may occur through the "Dexter" mechanism and the distance between these two kinds of pigments should be less than 1.nm. In addition, the results were also used to discuss the conformational relationship between p-Car and Chla molecules in CP43 and CP47.

关键词： CP43 CP47 β-Car Chla excitation energy transfer.

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体细胞克隆牛和转基因体细胞克隆牛的遗传学分析(英文)

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生物化学与生物物理进展 2004年第8期31卷 699-704页

作者：黄银花徐公瑾杜志强胡晓湘张磊涂剑锋程雪波李宁中国农业大学农业与生物技术国家重点实验室北京100094

分析了来自同一细胞系的体细胞克隆牛甜甜、庆庆、浒娃及来源同一培养转基因体细胞系转基因体细胞克隆牛九妹、乐娃和 1.妊娠 8个月转基因流产胎牛 8C2.及随机抽取的 1.鲁西黄牛 (LX)、 1.褐斯坦牛 (HS)在 2.4个微卫星位点标记牛的... 详细信息

分析了来自同一细胞系的体细胞克隆牛甜甜、庆庆、浒娃及来源同一培养转基因体细胞系转基因体细胞克隆牛九妹、乐娃和 1.妊娠 8个月转基因流产胎牛 8C2.及随机抽取的 1.鲁西黄牛 (LX)、 1.褐斯坦牛 (HS)在 2.4个微卫星位点标记牛的基因型 .结果表明 2.4个多态位点均表现出多态 ,等位基因数为 1. 5个 ,平均为 3 1.个 .根据网上公布的数据 ,按其最高频率计算 ,甜甜、庆庆、浒娃、九妹、乐娃、 8C2.培养细胞系、转基因细胞系间匹配概率为 1.1.× 1. -3 6,根据本研究观察到的数据计算 ,匹配概率为 1.90× 1. -2.3 ;而与随机抽取的 1.鲁西黄牛及褐斯坦牛的基因型分别在 2.3和2

关键词：体细胞克隆牛转基因遗传学等位基因

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Design and Expression of a Synthetic phyC Gene Encoding the Neutral Phytase in Pichia pastoris

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 803-811页

作者： li-Kou ZOU~(1,3) Hong-ning WANG~(2.1*) Xin PAN~4 Tao XIE~1 Qi WU~5 Zi-Wen XIE~1 Wan-Rong ZHOU~1 1 {2. of Veterinary and Biotechnology,Sichuan {2. university,Ya'an 62.014,china 2.Sichuan university Bioengineering Research Center for Animal Disease Prevention and Control,Chengdu 610065,china 3 Faculty of Environment and Resource of dujiangyan Campus,Sichuan {2. university,dujiangyan 611830,china 4 College of Earth Science,Chengd university of Technology,Chengdu 610059,china 5 College of life Science,Sichuan {2. university,Ya'an 62.014,china laboratory of Veterinary and Biotechnology Sichuan Agricultural University Ya'an 625014 China

The 1.74-bp phyCs gene （optimized phyC gene） encoding neutral phytase was designed andsynthesized according to the methylotrophic yeast Pichia pastoris codon usage bias without altering theprotein *** expression vector,pP9K-phyCs,was linearized and transformed in *** of total extracellular phytase activity was 1..6 U/ml in{1.ced in Buffered Methanol-complex Medium（BMMY） and 1..5 U/ml in Wheat Bran Extract In{1.ction （WBEI） medium at the flask scale,respectively,improving over 90 folds compared with the wild-type *** enzyme showed temperature optimumof 70℃ and pH optimum of *** enzyme activity retained 97% of the relative activity afterincubation at 80℃ for 5 *** of the heavy glycosylation the expressed phytase had a molecularsize of approximately 51.*** deglycosylation by endoglycosylase H （EndoH_f）,the enzyme had anapparent molecular size of 42.*** property and thermostability was affected by the glycosylation.

关键词： design expression synthetic phyC gene neutral phytase Pichia pastoris

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enOsCas1.0}1.mediated exon skipping for {1.chenne muscular dystrophy therapy in humanized mouse model

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Journal of Genetics and Genomics 2024年第2期51卷 256-259页

作者： Ming Jin Jiajia lin Yu Zhang Qingquan Xiao Xiangfeng Kong Xiumei Zhang Zhurui Shao Yin Wang Yuyang Yu Jinjing li Wan-Jin Chen Guoling li Hui Yang ning Wang Department of Neurology and Institute of Neurology of First Affiliated Hospital Institute of NeuroscienceFujian Key Laboratory of Molecular NeurologyFujian Medical UniversityFuzhouFujian 350004China HUIDAGENE Therapeutics Co. Ltd.Shanghai 200131China Institute of Neuroscience State Key Laboratory of NeuroscienceKey Laboratory of Primate NeurobiologyCAS Center for Excellence in Brain Science and Intelligence TechnologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

duchenne muscular dystrophy(DMD)is a severe neuromuscular disorder caused by mutations in the dystrophin gene,which encodes the essential protein *** genetic condition,affecting approximately 1.in 5000 male births worldwide(Birnkrant et al.,{1.1.),is characterized by progressive muscle weakness and wasting,leading to premature death in the second or third decade of life(Farini et al.,{1.1.).The absence of dystrophin protein causes instability in the sarcolemma,which predisposes individuals to myonecrosis and activation of inflammatory signaling ***,there is currently no effective cure for DMD,and glucocorticoid steroids are commonly used in clinical settings to delay symptom development(Merlini et al.,{1.03).

关键词： dystrophy duchenne

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The characteristics of CO_2.assimilation of photosynthesis and chlorophyll fluorescence in transgenic PEPC rice

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Chinese Science Bulletin 2001年第13期46卷 1080-1084页

作者： JIAO Demao, li Xia, HUANG Xueqing, CHI Wei, KUANG Tingyun & Ku SB Maurice1. Institute of Agrobiological Genetics and Physiology, Jiangsu Academy of {1. Sciences, Nangjing 2.001.,{2. 2. Institute of Botany, the Chinese Academy of Sciences, Beijing 1.0093,{2. 3. Botany Department, Washington state university, Pullman, WA 991.4-42.6, USA Institute of Agrobiological Genetics and Physiology Jiangsu Academy of Agricultural Sciences Nangjing China Institute of Botany The Chinese Academy of Sciences Beijing China Botany Department Washington State University Pullman USA

With PEPC, PPDK, NADP-ME and PEPC+ PPDK transgenic and untransformed rice (Orysa sativa L.), the activities of related C4 photosynthesis enzymes, the chlorophyll fluorescence parameters, CO2.exchange and other physiological indexes were compared, in which the physiological characteristics of PEPC transgenic rice were mainly studied. The results were as follows: (i) The activities of PEPC in PEPC transgenic rice were 2.-fold higher than those in untransformed rice; the light-saturation photosynthetic rates and the carboxylation efficiency of PEPC transgenic rice were increased by 55% and 50% more than those of untransformed rice, respectively, while the CO2.compensation point decreased by 2.%. (ii) The PS II photochemical efficiency (Fv/Fm) and photochemical quenching (qp) of transgenic PEPC rice decreased less in comparison with those of untransformed rice after the treatment with high light intensity (3 h) or methyl viologen (MV), a photooxidative reagent, which demonstrated that the tolerance of PEPC

关键词： transgenic rice photosynthetic characteristics photoin-hibition of photosynthesis chlorophyll fluorescence phosphoenopy-ruvate carboxylase (PEPC).

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CircMYBL2.faci{1.tates hepatocellular carcinoma progression by regulating E2.1.expression

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Oncology Research 2024年第6期32卷 1129-1139页

作者： JUNZHE YI BINBIN li XIAOMIN YIN liNGRUI liU CAILU SONG YING ZHAO MANBO CAI HAIliN TANG DONG CHEN ning LYU state {1. {2. of Oncology in South{2. Guangdong Provincial Clinical Research Center for CancerSun Yat-sen University Cancer CenterGuangzhou510060China Department of Medical Oncology The Third People’s Hospital of YongzhouYongzhou425000China Department of Radiotherapy The First Affiliated HospitalHengyang Medical SchoolUniversity of South ChinaHengyang421001China Center of Hepato-Pancreato-Biliary Surgery The First Affiliated HospitalSun Yat-sen UniversityGuangzhou510080China

Circular RNAs(circRNAs)have been recognized as pivotal regulators in tumorigenesis,yet the biological functions as well as molecular mechanisms of the majority of circRNAs in hepatocellular carcinoma(HCC)remain *** sought to unveil the expression profile and biological role of circMYBL2.in *** microarray analyses were con{1.cted to probe the expression profile of circMYBL2.in HCC cells,and qRT‒PCR analysis was then performed in HCC cell lines and tissues,revealing significant upregulation of *** experiments were con{1.cted to evaluate the biological function of circMYBL2.in HCC ***,bioinformatics analysis,qRT‒PCR analysis,luciferase reporter assays,and western blot analysis were employed to investigate the interplay among circMYBL2.miR-1.0}5,and ***2.was found to exhibit marked upregulation in tumor tissues as well as HCC cell *** expression of circMYBL2.increased the proliferation and migration of HCC cells,whereas circMYBL2.knockdown elicited contrasting ***,our results indicated that circMYBL2.promoted E2.1.expression and facilitated HCC progression by sponging *** findings revealed that circMYBL2.contributed to HCC progression through the circMYBL2.miR-1.0}5/E2.1.axis,suggesting the potential of circMYBL2.as a novel target for HCC treatment or a prognostic biomarker for HCC.

关键词： Circular RNAs circMYBL2 miR-1.0}5 E2.1 Hepatocellular carcinoma

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Effective accumulated temperature and developmental threshold temperature for Semanotus bifasciatus(Motschulsky) in Beijing

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Forest Ecosystems 2008年第2期10卷 125-129页

作者： MA li-qin1.GAO Shi-ji{1.WEN Jun-bao1.ZONG Shi-xiang1.XU Zhi-chun1.1.2} {{1. for Silviculture and Conservation of Ministry of Education,Beijing Forestry university,Beijing 1.0083,P.R.{2. {1.eijing agricultural Vocational College,Beijing 1.0088,P.R.{2. key {1. for Silviculture and Conservation of Ministry of Education Beijing Forestry University Beijing 100083 P. R. China Beijing agricultural Vocational College Beijing 100088 P. R. China

In order to understand the occurrence and development of Semanotus bifasciatus(Motschulsky),a borer insect of Platy-cladus orienta{1.s in parts of northern and eastern china and to improve its prediction,the developmen... 详细信息

In order to understand the occurrence and development of Semanotus bifasciatus(Motschulsky),a borer insect of Platy-cladus orientalis in parts of northern and eastern china and to improve its prediction,the developmental threshold temperature(Ct) and effective accumulated temperature(K) of *** were determined under conditions of constant and variable *** results show that under constant temperatures the value of Ct has a range of 8.90±1.77°C and the value of K lies between 95.19±13.14 degree-day for eggs;the values for larvae are Ct = 13.2.±3.06°C and the number of degree-day is K = 2.885.07±187.87 degree-day;for pupae the ranges are Ct = 8.93±2.49°C and K = 131.2.±2..63 *** the condition of ambient tempera-tures,the values for eggs are Ct = 17.33±1.2.°C and K = 70.79±8.99 *** is suggested that *** adults over-winter in a state of dormancy,not at a *** winter may accelerate the life cycle of *** can be used to predict an early occurrence of ***.

关键词： Semanotus bifasciatus(Motschulsky) developmental threshold temperature effective accumulated temperature eggs larvae pupae

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Numerical simulation of tidal current and erosion and sedimentation in the Yangshan deep-water harbor of Shanghai

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International Journal of Sediment Research 2009年第3期24卷 287-298页

作者： ZUO Shu-Hua1, ZHANG ning-Chuan2. li Bei3, ZHANG Zheng4, and ZHU Zhi-Xia51 Ph.D candidate, {2. key {2. of Coastal and Offshore Engineering, Dalian university of Technology, Dalian 11602., china key {2. of Engineering Sediment of Ministry of Communications, Tianjin Research Institute of Water Transport Engineering, Tianjin 300456, china. 2.Dr., Prof., {2. key {2. of Coastal and Offshore Engineering, Dalian university of Technology, Dalian 11602., china 3 Research Scientist, key {2. of Engineering Sediment of Ministry of Communications, Tianjin Research Institute of Water Transport Engineering, Tianjin 300456, china 4 Research Scientist, key {2. of Engineering Sediment of Ministry of Communications, Tianjin Research Institute of Water Transport Engineering, Tianjin 300456, china 5 Dr., Prof., Department of Harbor & Coastal Engineering, Shanghai Jiaotong university, Shanghai 2.0030, china [a] Ph.D candidate state {1. {2. of Coastal and Offshore Engineering Dalian {1. of Technology Dalian 1.60{1. china key {1. of Engineering Sediment of Ministry of Communications Tianjin Research Institute of Water Transport Engineering Tianjin 300456 china [b] Dr. Prof. state {1. {2. of Coastal and Offshore Engineering Dalian {1. of Technology Dalian 1.60{1. china [c] Research Scientist key {1. of Engineering Sediment of Ministry of Communications Tianjin Research Institute of Water Transport Engineering Tianjin 300456 china [d] Research Scientist key {1. of Engineering Sediment of Ministry of Communications Tianjin Research Institute of Water Transport Engineering Tianjin 300456 china [e] Dr. Prof. Department of Harbor & Coastal Engineering Shanghai Jiaotong university Shanghai 2.0030 china

Yangshan near-shore sea area is the multi-island and multi-channel area with strong flow velocity and high suspended sediment concentration. Based on the characteristics of tidal currents, waves, and sediment in the Yangshan area, a two-dimensional numerical model of tidal currents, sediment transport, and sea bed deformation is developed. In the model, the effects of tidal currents and wind waves on sediment transport are considered. According to characteristics of the study area, unstructured grids are applied to fit the boundaries of the near-shore sea area. The results show that the calculated values are in good agreement with the measured data. The field of tidal currents, suspended sediment concentrations, and the deformation of the seabed can be successfully simulated.

关键词： Yangshan deep-water port Tidal currents Sediment Suspended sediment concentration Seabed deformation Numerical simulation

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