ApoB RNA编辑高效蛋白检测体系的构建

[目的]建立一个载脂蛋白B(apoB)RNA编辑蛋白检测体系。[方法]在慢病毒表达质粒载体pCSII-CMV-IRES-Neor中插入apoB RNA编辑保守序列,并在其2端嵌入红色荧光蛋白(DsRed)、绿色荧光蛋白(GFP)表达序列,以此慢病载体转染大鼠肝癌细胞系CBRH-7919获得稳定表达。采用共聚焦显微镜测序方法检测apoB RNA的编辑。[结果]目的指示基因能够在CBRH-7919细胞中稳定表达,表现出指征RNA编辑的多色特征。[结论]试验成功构建了在细胞水平上以颜色变化指示apoB RNA编辑的检测体系,该体系为快速检测以apoB RNA编辑为靶的降胆固醇药物筛选提供了基础。     

Electron Transport Through Photosystem II Is Not Limited By A Wide Range of Water Deficit Conditions In Field‐Grown Gossypium hirsutum

Despite exhaustive literature describing drought stress effects on photosynthesis in Gossypium hirsutum, the sensitivity of photosynthetic electron flow to water deficit is heavily debated. To address this, G. hirsutum plants were grown at a field site near Camilla, GA under contrasting irrigation regimes, and pre‐dawn water potential (Ψ_PD), stomatal conductance (g_s), net photosynthesis (P_N), actual quantum yield of photosystem II (Φ_PSII) and electron transport rate (ETR) were measured at multiple times during the 2012 growing season. Ψ_PD values ranged from ?0.3 to ?1.1 MPa. Stomatal conductance exhibited a strong (r² = 0.697), sigmoidal response to Ψ_PD, where g_s was ≤0.1 mol m^?2 s^?1 at Ψ_PD values ≤ ?0.86 MPa. Neither Φ_PSII (r² = 0.015) nor ETR (r² = 0.010) was affected by Ψ_PD, despite exceptionally low Ψ_PD values (?1.1 MPa) causing a 71.7 % decline in P_N relative to values predicted for well‐watered G. hirsutum leaves at Ψ_PD = ?0.3 MPa. Further, P_N was strongly influenced by g_s, whereas ETR and Φ_PSII were not. We conclude that photosynthetic electron flow through photosystem II is insensitive to water deficit in field‐grown G. hirsutum.     

单细胞转录组测序技术及其在肌内脂肪细胞研究中的应用

单细胞转录组测序(single-cell RNA sequencing,scRNA-seq)技术在单细胞水平对转录组进行测序,可从表达量、细胞量及细胞组成等多种角度描述样本,主要包括单细胞分离、mRNA反转录、文库构建、转录组测序和数据分析等步骤。单细胞分离是scRNA-seq技术操作的第一步,常用的细胞分离方法有连续稀释法、显微操作法、荧光激活流式细胞术、激光捕获显微切割术和微流控技术。基于不同细胞捕获、cDNA扩增和文库构建开发了许多scRNA-seq方法,如CEL-seq2、Drop-seq、MARS-seq、SCRB-seq、Smart-seq、Smart-seq2等。与传统测序方法相比,scRNA-seq可识别单个细胞的基因表达信息、记录细胞的空间位置、跟踪不同细胞谱系在分化过程中的轨迹,这为研究难以大量提取的肌内脂肪细胞分子特异性和发生分化过程带来极大便利。作者简要介绍了scRNA-seq技术的关键步骤,比较分析了单细胞分离和转录组测序方法的优缺点,综述了肌内脂肪细胞的起源以及scRNA-seq在肌内脂肪细胞的亚群和标记基因鉴定以及细胞分化轨迹追踪中的应用,以期为肌内脂肪细胞的深入研究提供参考依据。     

二化螟dsRNA降解酶基因的克隆与表达分析

为解析二化螟Chilo suppressalis体内参与降解双链RNA(double-stranded RNA,dsRNA)的关键核酸酶的功能,克隆二化螟不同的非专一性核酸酶（non-specific nuclease,NUC）基因,并对这些基因进行生物信息学分析和组织定量表达分析,同时对dsRNA降解酶（dsRNA degrading nuclease,dsRNase）活力的组织分布进行研究。结果显示,共克隆获得5个NUC基因,其中有4个编码dsRNase亚家族基因（CsdsRNase1～CsdsRNase4）和1个编码Endonuclease G亚家族基因（CsEndoG）。5个NUC基因的开放阅读框核苷酸序列长度范围为828～1 338 bp,编码275～445个氨基酸残基,其分子量大小为31.68～49.57 kD,预测等电点为5.48～9.42。CsdsRNase1和CsdsRNase2含有信号肽序列,两者相似度极高,且与家蚕Bombyx mori和斜纹夜蛾Spodoptera litura中具有dsRNA降解酶活力的dsRNase同源聚类;CsdsRNase1和CsdsRN...     

抗流感病毒药物研究进展

季节性和爆发性流感引起了高发病率和死亡率,不仅严重危害人体健康,还影响社会稳定和经济发展。目前对抗流感的主要措施是预防接种,然而流感病毒变异速度非常快,使得现有的疫苗对新变异的流感病毒无效或者预防效果不好。因此,抗病毒药物的研究显得非常必要,而被深入研究的流感病毒的生命周期和病毒结构蛋白也推进了抗流感病毒药物设计的进展。本文将对抗流感药物设计靶向进行概述,旨在为今后研究开发出更高效、更理想的多靶向抗流感病毒药物提供借鉴。     

动物运输应激源与应激反应影响因子研究进展

动物的运输是畜牧业的重要生产环节,但运输活动带来的动物福利问题也越来越受到关注。开展动物运输应激源和应激反应影响因子的研究,对深入认识动物运输应激本质和提出解决方案乃至相关立法具有重要意义。论文对前人开展的动物运输应激源(拥挤、禁食禁水、气温、装卸、颠簸等)的分离研究和应激反应影响因子(运输距离和运输时间、年龄和经验、性情、恐惧信息素)的研究进行了回顾整理,并分析了争议的原因,为动物运输应激领域研究提供参考或指出新的研究方向。     

Bats as the main prey of wintering long‐eared owl (Asio otus) in Beijing: Integrating biodiversity protection and urban management

The loss of biodiversity from urbanized areas is a major environmental problem challenging policy‐makers throughout the world. Solutions to this problem are urgently required in China. We carried out a case study of wintering long‐eared owls (Asio otus) and their main prey to illustrate the negative effects of urbanization combined with ineffective conservation of biodiversity in Beijing. Field monitoring of owl numbers at two roosting sites from 2004 to 2012 showed that the owl population had fallen rapidly in metropolitan Beijing. Analysis of pellet contents identified only seven individuals of two species of shrew. The majority of mammalian prey comprised four bat and seven rodent species, making up 29.3% and 29.5% of the prey items, respectively. Prey composition varied significantly among years at the two sample sites. At the urban site the consumption of bats and rodents declined gradually over time, while predation on birds increased. In contrast, at the suburban site the prey composition showed an overall decrease in the number of bats, a sharp increase and a subsequent decrease in bird prey, and the number of rodent prey fell to a low point. Rapid development of real estate and inadequate greenfield management in city parks resulted in negative effects on the bird and small mammal habitat of urban areas in Beijing. We suggest that measures to conserve biodiversity should be integrated into future urban planning to maintain China's rich biodiversity while also achieving sustainable economic development.