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Recombinant Expression of the Monomeric B27 Lys Destripeptide Insulin Precursor( MIP) in Escherichia coli

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Journal of Donghua University(English Edition) 2018年第1期35卷 43-46页

作者： CHEN Ting ZHU Wenkai ZHANG Yunlong ZHANG Minmin CHEN Li lu changrui College of Chemistry Chemical Engineering and Biotechnology Donghua University Shanghai 201620 China

Insulin controls hyperglycemia caused by diabetes,and virtually all treatments require exogenous insulin. The monomeric B27 Lys destripeptide insulin has 80% bioactivity of wild type. It is a potential drug for clinic. But the yield of monomeric B27 Lys destripeptide insulin is limited, because monomeric B27 Lys destripeptide insulin precursor( MIP) was easily to form inclusion body,when MIP is expressed in E. coli. The precursor,MIP,is not only fused to an N-terminal HIS-small ubiquitin-related modifier( SUMO) tag,but also added five Arginine( 5 R) at the C-terminal,which can improve the solubility of the fusion protein. By this way,the yield of fusion protein has been up to 1. 45 mg/L cell *** work set up the foundation for the clinical application of B27 Lys destripeptide insulin.

关键词： B27 Lys destripeptide insulin optimization monomeric B27 Lys destripeptide insulin precursor(MIP)

Raoultella terrigena RtZG1 Electrical Performance Appraisal and System Optimization

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Journal of Donghua University(English Edition) 2021年第3期38卷 252-256页

作者： GONG Kang CHEN Ting LÜ Liangyu BAO Guangsen lu changrui ZHANG Yunlong College of Chemistry Chemical Engineering and Biotechnology Donghua University Shanghai 201620 China

The aims of this study are to provide a strain and its optimal application in electro-producting performance for revealing the electrical generating mechanism in the *** constructed microbial fuel cells(MFCs)device that was a bipolar chamber MFC,using metal ion media(MIM)and water as anode and cathode reaction substation fluid,*** this study,we identified an isolate as Raoultella terrigena,named RtZG1,which could produce ***,we optimized the conditions of electrical energy *** continuous output current could reach about 200μA within 3 h when the ratio of electro-bacterial fluid to matrix fluid was 1∶4,the temperature was 37℃,the carbon-nitrogen ratio of the inorganic salt medium was 10∶1,as well as the concentration of MIM was *** on the optimization,it is clear that the most suitable conditions of electricity production for this strain lay the foundation for the application of this strain.

关键词： microbial fuel cell(MFC) Raoultella terrigena electro-bacteria conditions optimization

Optimization of RNA Secondary Structure Prediction with SHAPE(Selective 2'-hydroxyl Acylation Analyzed by Primer Extension)Chemistry

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Agricultural Biotechnology 2017年第1期6卷 52-54页

作者： Jun ZHENG Rujie CAI Altaf SIMAIR Ting CHEN Yunlong ZHANG changrui lu College of Chemistry Chemical Engineering and BioengineeringDonghua UniversityShanghai 201620China

SHAPE （selective T-hydroxyl acylation analyzed by primer extension） chemistry has been widely used in the prediction of RNA secondary structure at single-nucleotide solution. This study was designed to determine the optimal concentrations of RNA and modifying reagent in SHAPE analysis to improve the accura- cy of structure prediction. The results showed that at the ratio of N-methylisatuic anhydride （NMIA）（μmol/L） to RNA （0μmol/L） of 13： 20, the resulting concen- tration of full-length RNA was the highest, and the sequencing signal was the strongest. So, this ratio is optimal for the analysis and prediction of RNA secondary structure in SHAPE chemistry.

关键词： SHAPE RNA concentration NMIA concentration Optimization

Expression and Purification of Zinc Finger Domain and Central Domain of MDMD2

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Agricultural Biotechnology 2016年第2期5卷 37-39页

作者： Xiaoling REN Ting CHEN Yunlong ZHANG changrui lu Minmin ZHANG Jun ZHENG Xiaoqi QI Donghua University Shanghai 201600 China

[ Objective] This study aimed to construct the His-tagged prokaryotic expression vectors harboring zinc finger domain （ZF） and central domain （ acidic domain ＆ zinc finger domain, CT） of MDM2, respectively, and preliminarily identified biologic activity of the purified fusion proteins. [ Method ] ZF and CT cod- ing regions were amplified from MDM2 cDNA library by PCR and separately inserted into prokaryotie expression vector pET28b to construct the recombinant plas- mids. After verification by enzyme digestion, the recombinant plasmids were separately transformed into E. coli DH5c~ competent cells. The expressed recombinant plasmids were purified using Ni-NTA magnetic beads and identified by SDS-PAGE and Western blotting. [ Result] The molecular weight of His-ZF and His-CT fu- sion proteins was 21 and 31 kD, respectively. E Conclusion] Recombinant fusion proteins containing ZF and CT of MDM2 were obtained successfully, which laid the foundation for subsequent protein crystallization and three-dimensional structure analysis.

关键词： Zincfinger domain Central domain MDM2 Prokaryotic expression Protein purification

Prediction of the Structure of Full-length Human Telomer-ase RNA with SHAPE(Selective 2'-hydroxyl Acylation An-alyzed by Primer Extension)

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Agricultural Biotechnology 2017年第1期6卷 55-57,60页

作者： Xiaoqi QI Yunlong ZHANG Ting CHEN Altaf Ahmed SIMAIR Minmin ZHANG Zhuoru YANG changrui lu College of Chemistry Chemical Engineering and BioengineeringDonghua UniversityShanghai 201620China

Telomerase is a unique ribonucleoprotein complex that protects chromosomes from degradation and recombination. The defects in telomerase activity eansed by the mutations in telomerase RNA （hTER） is associated with several human genetic diseases, so the molecular biology and structural biology of hTER are of great significance for the treatment of related diseases and the development of corresponding drugs. In this study, hTER was obtained by in vitro transcription, and then modified with N-methylisatoic anhydride （NMIA） by SHAPE （selective 2＇-hydroxyl aeylation analyzed by primer extension） ; finally, the eDNA fragment was obtained by reverse transcription to analyze the free nucleotides and dynamic structure of hTER, which may provide experimental basis for analyzing three--di- mensional structure of hTER obtained in vitro. In addition, the SHAPE parameters for long RNA fragment （〉 300 nt） were optimized. The results showed that 1 μmol/L hTER and 3.9 mmol/L NMIA were optimal, and 49 ℃ was the best temperature for primer extension.

关键词： HTER In vitro transcripton RNA purification SHAPE experiment

Small Amplicons Mutation Library for Vaccine Screening by Error-Prone Polymerase Chain Reaction

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Journal of Donghua University(English Edition) 2023年第2期40卷 171-176页

作者：程曼曼张云龙陈婷张敏敏陆昌瑞 College of Chemistry Chemical Engineering and BiotechnologyDonghua UniversityShanghai 201620China

Library construction is a common method used to screen target genes in molecular *** library constructions are not suitable for a small DNA library(<100 base pair(bp))and low RNA library *** maximize the library’s co... 详细信息

Library construction is a common method used to screen target genes in molecular *** library constructions are not suitable for a small DNA library(lution and amplification of error-prone *** error-prone PCR conditions were optimized including deoxyribonucleotide triphosphate(dNTP)concentration,Mn^(2+)concentration,Mg^(2+)concentration,PCR cycle number,and primer ***,a RNA library with high complexity can be obtained by in vitro transcription to meet most molecular biological screening requirements,and can also be used for mRNA vaccine screening.

关键词： error-prone polymerase chain reaction in vitro transcription DNA library RNA library

跨境社会资本对跨境微区域合作的影响路径分析

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云南行政学院学报 2023年第4期25卷 1-14页

作者：曹亚斌卢昌瑞西北师范大学西北边疆政治与治理研究中心甘肃兰州730070 西北师范大学马克思主义学院甘肃兰州730070

较之于宏观层次的区域合作,作为区域合作微观形式的跨境微区域合作具有空间范围小、投入资金少、地方性行为体的作用更为突出、对国家间关系的整体影响小等特性,这使得在跨境微区域合作中跨境社会资本往往能发挥更为重要的作用。跨境社... 详细信息

较之于宏观层次的区域合作,作为区域合作微观形式的跨境微区域合作具有空间范围小、投入资金少、地方性行为体的作用更为突出、对国家间关系的整体影响小等特性,这使得在跨境微区域合作中跨境社会资本往往能发挥更为重要的作用。跨境社会资本对跨境微区域合作的影响主要通过其所具有的密度、关键节点中心势两条路径予以实现:一方面,随着跨境社会资本密度的不断增大,各跨境行为体间的交易成本随之越低,跨境微区域合作成功的概率增大;另一方面,随着跨境社会资本关键节点中心势的不断集中,集体行动困境更容易被克服,跨境微区域合作成功的概率也会增大。本文运用Cox比例风险模型对上述影响路径进行了检验,证明了跨境社会资本对于跨境微区域合作存在着显著的正向影响力。

关键词：跨境社会资本跨境微区域合作 Cox回归分析

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局部阴影最大功率点MPPT算法研究

电力科学与工程 2012年第5期28卷 12-15页

作者：吕昌睿李国杰电力传输与功率变换控制教育部重点实验室上海交通大学电气工程系上海200240

光伏阵列作为太阳能光伏发电系统的基本发电单元,容易受到阴影的影响。在局部阴影条件下,光伏阵列的输出特性发生改变,相应的功率电压曲线含有多个局域峰值,使常规的最大功率点跟踪算法很难准确地跟踪到真正的最大功率点。在光伏电池通... 详细信息

光伏阵列作为太阳能光伏发电系统的基本发电单元,容易受到阴影的影响。在局部阴影条件下,光伏阵列的输出特性发生改变,相应的功率电压曲线含有多个局域峰值,使常规的最大功率点跟踪算法很难准确地跟踪到真正的最大功率点。在光伏电池通用数学模型的基础上,结合串并联理论,对局部阴影条件下光伏阵列的输出特性进行了数学建模分析,结果表明,带检测环节的MPPT算法有更好的适应性和稳定性。

关键词：光伏阵列输出特性局部阴影最大功率点跟踪

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应用微机程序进行铸件废品统计分析

热加工工艺 1988年第6期 25-27页

作者：叶荣茂陆发天祝长瑞哈尔滨工业大学哈尔滨汽轮机厂

本文介绍了应用微机程序进行废品统计分析的方法。生产实际应用表明,采用微机程序进行铸件废品统计分析能够快速、准确地提供各种废品统计、分析图表及统计数据。程序运行操作方便,不要求操作人员具有计算机方面的知识。

关键词：铸件废品统计分析财政管理铸造生产图号