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11.
Zhengquan Yang Chengliang Zhang Guojun Lian Shijie Dong Menghui Song Hengrong Shao Jingmei Wang Tao Zhong Zhenni Luo Shengnan Jin Chunming Ding 《Nucleic acids research》2022,50(13):7560
5′-Adenylated oligonucleotides (AppOligos) are widely used for single-stranded DNA/RNA ligation in next-generation sequencing (NGS) applications such as microRNA (miRNA) profiling. The ligation between an AppOligo adapter and target molecules (such as miRNA) no longer requires ATP, thereby minimizing potential self-ligations and simplifying library preparation procedures. AppOligos can be produced by chemical synthesis or enzymatic modification. However, adenylation via chemical synthesis is inefficient and expensive, while enzymatic modification requires pre-phosphorylated substrate and additional purification. Here we cloned and characterized the Pfu RNA ligase encoded by the PF0353 gene in the hyperthermophilic archaea Pyrococcus furiosus. We further engineered fusion enzymes containing both Pfu RNA ligase and T4 polynucleotide kinase. One fusion enzyme, 8H-AP, was thermostable and can directly catalyze 5′-OH-terminated DNA substrates to adenylated products. The newly discovered Pfu RNA ligase and the engineered fusion enzyme may be useful tools for applications using AppOligos. 相似文献
12.
Yu Shang Li Li Tengfei Zhang Qingping Luo Qingzhong Yu Zhe Zeng Lintao Li Miaomiao Jia Guoyi Tang Sanlin Fan Qin Lu Wenting Zhang Yuhan Xue Hongling Wang Wei Liu Hongcai Wang Rongrong Zhang Chan Ding Huabin Shao Guoyuan Wen 《PLoS pathogens》2022,18(6)
The development of thermostable vaccines can relieve the bottleneck of existing vaccines caused by thermal instability and subsequent poor efficacy, which is one of the predominant reasons for the millions of deaths caused by vaccine-preventable diseases. Research into the mechanism of viral thermostability may provide strategies for developing thermostable vaccines. Using Newcastle disease virus (NDV) as model, we identified the negative surface charge of attachment glycoprotein as a novel determinant of viral thermostability. It prevented the temperature-induced aggregation of glycoprotein and subsequent detachment from virion surface. Then structural stability of virion surface was improved and virus could bind to and infect cells efficiently after heat-treatment. Employing the approach of surface charge engineering, thermal stability of NDV and influenza A virus (IAV) vaccines was successfully improved. The increase in the level of vaccine thermal stability was determined by the value-added in the negative surface charge of the attachment glycoprotein. The engineered live and inactivated vaccines could be used efficiently after storage at 37°C for at least 10 and 60 days, respectively. Thus, our results revealed a novel surface-charge-mediated link between HN protein and NDV thermostability, which could be used to design thermal stable NDV and IAV vaccines rationally. 相似文献
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Ennian Li Kai Wang Bei Zhang Siqi Guo Senhao Xiao Qi Pan Xiaowan Wang Weiying Chen Yunshan Wu Hesong Xu Xiangqian Kong Cheng Luo Shijie Chen Bo Liu 《Journal of enzyme inhibition and medicinal chemistry》2022,37(1):1537
The DNA methyltransferases (DNMTs) were found in mammals to maintain DNA methylation. Among them, DNMT1 was the first identified, and it is an attractive target for tumour chemotherapy. DC_05 and DC_517 have been reported in our previous work, which is non-nucleoside DNMT1 inhibitor with low micromolar IC50 values and significant selectivity towards other S-adenosyl-L-methionine (SAM)-dependent protein methyltransferases. In this study, through a process of similarity-based analog searching, a series of DNMT1 inhibitors were designed, synthesized, and evaluated as anticancer agents. SAR studies were conducted based on enzymatic assays. And most of the compounds showed strong inhibitory activity on human DNMT1, especially WK-23 displayed a good inhibitory effect on human DNMT1 with an IC50 value of 5.0 µM. Importantly, the pharmacokinetic (PK) profile of WK-23 was obtained with quite satisfying oral bioavailability and elimination half-life. Taken together, WK-23 is worth developing as DNMT1-selective therapy for the treatment of malignant tumour. 相似文献
15.
Lifang Guo Xingchao Geng Li Liu Yufa Miao Zhi Lin Min Yu Yan Fu Lihong Liu Bo Li Yongzhang Luo 《Journal of biochemical and molecular toxicology》2019,33(3)
Endostar, a potent endogenous antiangiogenic factor, is wildly used in clinics. However, it was easily degraded by enzymes and rapidly cleared by the kidneys. To overcome these shortcomings, PEGylated recombinant human endostatin was developed. In this study, the purity of M2ES was evaluated by silver stain and reversed‐phase high‐performance liquid chromatography. Ultraviolet spectrum was used to examine the structural of M2ES and endostar. The bioactivity and antitumor efficacy of M2ES were evaluated using an in vitro endothelial cell migration model and athymic nude mouse xenograft model of a heterogeneous lung adenocarcinoma, respectively. A preclinical study was performed to evaluate the acute toxicity and safety pharmacology in rhesus monkeys. The purity of M2ES was more than 98%; PEG modification has no effect on endostatin structure. Compared with the control group, M2ES dramatically retards endothelial cell migration and tumor growth. After intravenous (IV) infusions of M2ES at a dose level of three and 75 mg/kg in rhesus monkeys, there was no observable serious adverse event in both acute toxicity and safety pharmacology study. On the basis of the quality and bioactivity study data of M2ES and the absence of serious side effect in rhesus monkeys, M2ES was authorized to initiate a phase I clinical trial. 相似文献
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Junna Wang Chong Li Xinbin Duan Daqing Chen Shunxin Feng Huihuang Luo Qidong Peng Wengen Liao 《Freshwater Biology》2014,59(7):1343-1360
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18.
贺兰山马鹿冬春季生境的选择 总被引:4,自引:2,他引:4
2007年12月~2008年1月和2008年4~5月,在贺兰山利用痕迹检验法和直接观察法对马鹿阿拉善亚种冬春季生境选择进行研究.通过在选定的15条沟段里进行调查,冬春季各测定了131个和181个样方的18种生态因子.结果表明,冬春季贺兰山马鹿在植被类型、优势乔木、灌木密度、灌木高度、灌木距离、坡向、坡位、海拔高度、距裸岩距离和隐蔽级10种生态因子上差异极显著(P < 0.001),其余生态因子无显著差异(P < 0.05),相对于冬季生境而言,马鹿春季更偏爱选择以油松和青海云杉为优势乔木的山地针叶林带,灌木密度大、高度高、距离近,半阴半阳坡的中坡位,海拔较高,距裸岩较近,隐蔽程度高的生境.典则判别系数显示冬春季马鹿在生境选择上存在一定程度的重叠,但春季马鹿的分布范围要比冬季更广一些,Wilk's λ值显示冬春季马鹿的生境选择具有较高的差异性(Wilk's λ = 0.683,x2 = 116.995,df = 13,P < 0.001).逐步判别分析表明在区分冬春季马鹿生境方面有一系列生态因子发挥作用,依照贡献值的大小依次为:乔木密度、灌木距离、坡度、海拔高度、距水源距离、距裸岩距离和隐蔽级,由这7个变量构成的方程对冬春季马鹿生境的正确区分率达到79.9%,其中对春季的误判率为22.1%,对冬季的误判率为20.6%. 相似文献
19.
甘肃矿产资源丰富,矿区酸性矿坑水(acid mine drainage,AMD)及周边土壤中嗜酸微生物种类丰富,具有相当大的研究价值.本研究通过Illumina高通量测序方法,对甘肃省4个矿区AMD及周边土壤中的细菌群落组成、相似性及功能组成进行分析.结果 表明,煤矿样品GNM细菌物种组成最为丰富,金矿样品LNJ1与紫金矿样品LNZJK细菌组成最相似,与属于铜矿的BYT物种较相近,与LNJ1同地区的LNJ2样品细菌组成相比较远.所有样品细菌主要分为放线菌门(Actinobacteria)、拟杆菌门(Bacteriodetes)、厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)四大门,其中变形菌门(Proteobacteria)丰度最高.放线菌门(Actinobacteria)中不动杆菌属(Acinetobacter)、类诺卡氏属(Nocardioides)和芽球菌属(Blastococcus)表现出较高丰度;厚壁菌门(Firmicutes)中微小杆菌属(Exiguobacterium)和芽孢杆菌属(Bacillus)表现出较高丰度,且微小杆菌属(Exiguobacterium)丰度极高.通过功能预测发现,与煤矿样品GNM相比,LNJ1、LNJ2、BYT和LNZJK4个金属矿区样品中的微生物具有较强的氨基酸与碳源的转运与代谢、转录等功能以及较低的能量生产与转运、无机离子转运与代谢能力.通过探究AMD中的细菌多样性及其与重金属之间的相互关系,了解AMD采样区微生物群落结构,以期挖掘出更为丰富的具有重金属抗性的菌株资源,以应用于生物浸矿和环境修复等领域. 相似文献
20.
Hui Luo Jing Xiong Qiaoni Zhou Liqiu Xia Ziquan Yu 《Applied microbiology and biotechnology》2013,97(23):10135-10142
Several families of crystal proteins from Bacillus thuringiensis exhibit nematicidal activity. Cry5B protein, a pore-forming toxin, has been intensively studied yielding many insights into the mode of action of crystal protein at molecular level and pathogenesis of pore-forming toxins. However, little attention was paid to Cry6A, another representative nematicidal crystal protein. Cry6A shares very low homology with Cry5B at amino acid sequence and probably acts in a distinct pathway from Cry5B and even the other main commercial crystal proteins. In the current study, we comprehensively investigated the nematicidal properties of Cry6Aa2 against the free-living soil nematode Caenorhabditis elegans and examined the physical response of C. elegans to Cry6Aa2 attack. Our results indicate that Cry6Aa2 exhibits high lethal activity to C. elegans and could cause detrimental effects on C. elegans, including obviously suppressed growth, decreased brood size, and even abnormal motility. Meanwhile, our study additionally shows that C. elegans could defend against the Cry6Aa2 toxin harmful threat through behavioral defense responses, such as reduced oral uptake and physical avoidance. In general, this study suggests that Cry6Aa2 possesses diverse nematicidal properties, which strongly indicates that Cry6Aa2 is a promising potential candidate of nematicidal agent. Moreover, this study highlights the importance of behavioral responses in defense of C. elegans for survival and demonstrates the key role of crystal protein in the interaction of B. thuringiensis–C. elegans. These findings could shed light on understanding the interaction of C. elegans with B. thuringiensis and provide a perfect model to study the role of pathogenic factor in the interaction of pathogen–host. 相似文献