Direct adenylation from 5′-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase

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.     

三种杀虫菌超氧化物歧化酶电泳图型和同源性

本文报道了球形芽孢杆菌、苏云金芽孢杆菌和森田芽孢杆菌三种昆虫病原菌的超氧化物歧化酶（SOD）的电泳图型及其同源性。实验证明B．S和B．tSOD分别具有Ef25～30、56．8和52．9特征酶带,B．mSOD也具有与B．T相同的特征酶带;免疫琼脂双扩散实验证明B．sSOD与B．T和B．mSOD无同源性,B．tSOD抗血清与B.sSOD抗原无沉淀反应,但与B．mSOD抗原产生沉淀反应,证明B．S与B.t和B．M的亲缘关系远,B．T和B．M的亲缘关系近。     

球形芽孢杆菌C3-41超氧化物岐化酶的特性

本文研究了球形芽孢杆菌（Bacillussphaericus）C3－41超氧化物歧化酶（SOD）的产生条件和部分特性。当C3－41菌株处于孢子囊中期时为产SOD酶高峰期,在30℃下的平板培养物及培养基起始pH为中性（pH7．0）时产生的SOD酶比活最高,经硫酸铵分级沉淀,DEAE－32离子交换层析和SephadexG-100凝胶过滤提纯了SOD酶。此酶属Mn－SOD,在25～35℃和pH5～9范围内较稳定,但在55℃下10min完全失活。     

曲酸菌选育及发酵工艺研究

筛选获得产曲酸菌株米曲霉（Aspergillusoryzae）MSA进行⁶⁰Co诱变,并进行了发酵工艺条件研究,以葡萄糖为碳源、豆饼粉为氮源,在pH3.0、33℃摇瓶培养,可达到5d产酸5％以上水平。发酵液采用直接浓缩结晶工艺,脱色与重结晶后获得无色针状晶体,红外图谱检测确证为曲酸。     

Distinct basolateral amygdala excitatory inputs mediate the somatosensory and aversive-affective components of pain

Pain is a multidimensional perception that includes unpleasant somatosensory and affective experiences; however, the underlying neural circuits that mediate different components of pain remain elusive. Although hyperactivity of basolateral amygdala glutamatergic (BLA^Glu) neurons is required for the somatosensory and emotional processing of pain, the precise excitatory inputs to BLA^Glu neurons and their roles in mediating different aspects of pain are unclear. Here, we identified two discrete glutamatergic neuronal circuits in male mice: a projection from the insular cortex glutamatergic (IC^Glu) to BLA^Glu neurons, which modulates both the somatosensory and affective components of pain, and a projection from the mediodorsal thalamic nucleus (MD^Glu) to BLA^Glu neurons, which modulates only the aversive-affective component of pain. Using whole-cell recording and fiber photometry, we found that neurons within the IC→BLA and MD→BLA pathways were activated in mice upon inflammatory pain induced by injection of complete Freund’s adjuvant (CFA) into their paws. Optical inhibition of the IC^Glu→BLA pathway increased the nociceptive threshold and induced behavioral place preference in CFA mice. In contrast, optical inhibition of the MD^Glu→BLA pathway did not affect the nociceptive threshold but still induced place preference in CFA mice. In normal mice, optical activation of the IC^Glu→BLA pathway decreased the nociceptive threshold and induced place aversion, while optical activation of the MD^Glu→BLA pathway only evoked aversion. Taken together, our results demonstrate that discrete IC^Glu→BLA and MD^Glu→BLA pathways are involved in modulating different components of pain, provide insights into its circuit basis, and better our understanding of pain perception.     

基于遥感降尺度估算中国森林生物量的空间分布

森林生物量是陆地生态系统重要的碳库,其大小与空间分布特征直接影响森林的碳汇潜力。基于空间降尺度技术,以中国第六次国家森林资源清查资料为基础,同时结合1∶100万植被分布图及同期的基于MODIS反演的NPP空间分布,定量估算了1 km分辨率下我国森林生物量的空间分布。结果表明:(1)降尺度技术能有效结合遥感数据的空间特征与地面详查资料的统计特征,从而较好地解决当前生物量估算的区域尺度转化问题;(2)我国森林生物量存在明显的空间分布规律,与水热条件的空间分布格局基本一致,表现为西部较低东部较高,大型山脉分布处较高;(3)我国森林生物量总量11.0 Pg,平均生物量74.8 Mg/hm2,其中高值区主要集中在东北大小兴安岭和长白山地区、新疆山区、西南横断山脉地区以及东南武夷山地区。     

瘦蛋白与皮肤创伤愈合

自1994年瘦蛋白（leptin）被发现以来,其生理作用得到广泛的研究。近年的研究表明,在高等动物的皮肤创伤愈合过程中,胶原与血管的生成、肉芽组织与上皮的再生以及组织的重建等因子,对皮肤的创伤愈合发挥着至关重要的作用,了解瘦蛋白与上述因子之间的相互关系与作用机制,有助于指导治疗,改善预后。