CRISPR/Cas基因编辑系统在原核微生物细胞工厂构建中的开发与应用

随着能源和环境问题的日益突出,化学品以及燃料的合成方式正逐渐由传统的化学法合成转变为以细菌为基础的生物炼制过程,其中最关键问题是需要开发出合适的基因工程工具用于构建相应的产品生产菌株。成簇的规律间隔短回文重复序列(Clusteredregularlyinterspacedshortpalindromic repeats,CRISPR)/CRISPR相关蛋白(CRISPR-associated proteins,Cas)系统是一种存在于细菌和古细菌中的免疫系统,能够用于抵御病毒和外源质粒的入侵,近年来被开发成为一种高效、便捷、精确的基因编辑工具,显示出巨大的应用潜力。本文立足于CRISPR/Cas系统的原理与最新分类,结合实例综述了CRISPR/Cas基因编辑系统在原核微生物细胞工厂构建中的建立与优化策略,以及主要的应用方向,并探讨该系统所面临的主要问题并提出了一些可行的解决方案。     

Epidemiological Characteristics of Influenza A and B in Macau, 2010–2018

Virologica Sinica - Influenza is one of the major respiratory diseases in humans. Macau is a tourist city with high density of population and special population mobility. The study on the...     

Effect of surgical castration on expression of TRPM8 in urogenital tract of male rats

Trpm8 (melastatin-related transient receptor potential member 8), a member of the transient receptor potential (TRP) superfamily, encoding a cation channel named TRPM8, has been shown to be a primary androgen-responsive gene and play an important role in prostate physiology. To investigate the expression feature of TRPM8 in urogenital tract of male rats, and whether TRPM8 was also regulated by androgen receptor in these organs, male Sprague–Dawley rats were divided into three groups of 35 animals as follows: sham-operated (SHAM), orchidectomized (ORX), orchidectomized plus DHT treatment (O + D). Organs in urogenital tract, including kidney, prostate, seminal vesicle (SV), testis, epididymis and penis, were collected at different post-castration periods. RT-PCR, real-time PCR and Western blotting were used to detect the expression of androgen receptor (AR) and trpm8 in these tissue. As a result, AR and trpm8 can be detected in all these organs at mRNA or/and protein level. The mRNA expression of trpm8 in kidney, prostate, SV and penis decreased 24 or 72 h after castration and kept decreasing in a time-dependant manner. However, treatment of dihydrotestosterone (DHT) could reverse the effect of surgical castration. Collectively, our data provide evidence that TRPM8 and AR were expressed generally in urogenital tract of male rats, and in these organs, expression of trpm8 was regulated by serum androgen.     

2008-2014年新疆艾比湖流域土壤水分时空分布特征

传统的土壤水分模拟研究难以从土壤水分变化的时空双向出发表达其连续演变的过程,存在时空尺度效应问题。借助SWAT模型模拟的长时间序列优势,结合高分辨率卫星影像和遥感技术,力图在时空尺度效应问题上取得突破。并利用长时间序列的模拟结果分析流域土壤水分的空间格局和不同维度时空异质性。结果表明：（1）2008-2014年间艾比湖流域土壤水分主要受气温、降水及人类活动影响,呈波动变化,总体偏低且具有逐年减小趋势。（2）受降水、地形及土地覆被影响,土壤水分分布呈现出由山区向两侧平原减少的特点,且林地 > 农用地 > 草地 > 稀疏植被。（3）近10年间土壤水分低值区由原来的北部山区及平原向东部、东南部平原区及南部山区迁移,东部减少最为明显。（4）流域四季土壤水分变化差异显著。其中,春季主要受融雪影响;夏季、秋季主要受降雨量和气温影响;冬季主要受固态降雪和气温影响;且不同年份、相同季节、相同子流域土壤水分变化趋势表现一致。     

大草蛉对草地贪夜蛾捕食潜能研究

草地贪夜蛾Spodoptera frugiperda JE Smith于2018年底至2019年初入侵我国后,迅速扩散,并在半年内对西南地区的玉米生产造成为害。田间调查和观测研究表明,田间虽然可见该害虫的天敌,但由于天敌种群数量低,无法形成有效控害的模式。为研究大草蛉对草地贪夜蛾的控害潜能,本研究进行了捕食功能反应试验。结果表明,大草蛉成虫对草地贪夜蛾卵以及大草蛉3龄幼虫对草地贪夜蛾1龄幼虫的捕食效应均能够很好的拟合HollingⅡ功能反应模型。其中,大草蛉成虫对贪夜蛾卵的理论日最大捕食量、瞬时攻击率和处理时间分别为1115.56头、1.004和0.0009 d。大草蛉幼虫对贪夜蛾低龄幼虫的理论日最大捕食量、瞬时攻击率和处理时间分别为358头、1.074和0.003 d。结果表明大草蛉具备对草地贪夜蛾卵和低龄幼虫进行有效控害的潜力,为应用天敌昆虫防治草地贪夜蛾提供了理论依据。     

Analysis of metabolic and physiological responses to gnd knockout in Escherichia coli by using C-13 tracer experiment and enzyme activity measurement

The physiological and metabolic responses to gnd knockout in Escherichia coli K-12 was quantitatively investigated by using the (13)C tracer experiment (GC-MS/NMR) together with the enzyme activity analysis. It was shown that the general response to the gene knockout was the local flux rerouting via Entner-Doudoroff pathway and the direction reversing via non-oxidative pentose phosphate pathway (PPP). The mutant was found to direct higher flux to phosphoglucose isomerase reaction as compared to the wild-type, but the respiratory metabolism was comparable in both strains. The anaplerotic pathway catalyzed by malic enzyme was identified in the mutant, which was accompanied with an up-regulation of phosphoenolpyruvate carboxylase and down-regulation of phosphoenolpyruvate carboxykinase. The presented results provide first evidence that compensatory mechanism existed in PPP and anaplerotic pathway in response to the gnd deletion.     

Genome sequences of siphoviruses infecting marine Synechococcus unveil a diverse cyanophage group and extensive phage-host genetic exchanges

Investigating the interactions between marine cyanobacteria and their viruses (phages) is important towards understanding the dynamic of ocean's primary productivity. Genome sequencing of marine cyanophages has greatly advanced our understanding about their ecology and evolution. Among 24 reported genomes of cyanophages that infect marine picocyanobacteria, 17 are from cyanomyoviruses and six from cyanopodoviruses, and only one from cyanosiphovirus (Prochlorococcus phage P-SS2). Here we present four complete genome sequences of siphoviruses (S-CBS1, S-CBS2, S-CBS3 and S-CBS4) that infect four different marine Synechococcus strains. Three distinct subtypes were recognized among the five known marine siphoviruses (including P-SS2) in terms of morphology, genome architecture, gene content and sequence similarity. Our study revealed that cyanosiphoviruses are genetically diverse with polyphyletic origin. No core genes were found across these five cyanosiphovirus genomes, and this is in contrast to the fact that many core genes have been found in cyanomyovirus or cyanopodovirus genomes. Interestingly, genes encoding three structural proteins and a lysozyme of S-CBS1 and S-CBS3 showed homology to a prophage-like genetic element in two freshwater Synechococcus elongatus genomes. Re-annotation of the prophage-like genomic region suggests that S.?elongatus may contain an intact prophage. Cyanosiphovirus genes involved in DNA metabolism and replication share high sequence homology with those in cyanobacteria, and further phylogenetic analysis based on these genes suggests that ancient and selective genetic exchanges occurred, possibly due to past prophage integration. Metagenomic analysis based on the Global Ocean Sampling database showed that cyanosiphoviruses are present in relatively low abundance in the ocean surface water compared to cyanomyoviruses and cyanopodoviruses.     

Highly luminescent S,N co‐doped carbon quantum dots‐sensitized chemiluminescence on luminol–H2O2 system for the determination of ranitidine

S,N co‐doped carbon quantum dots (N,S‐CQDs) with super high quantum yield (79%) were prepared by the hydrothermal method and characterized by transmission electron microscopy, photoluminescence, UV–Vis spectroscopy and Fourier transformed infrared spectroscopy. N,S‐CQDs can enhance the chemiluminescence intensity of a luminol–H₂O₂ system. The possible mechanism of the luminol–H₂O₂–(N,S‐CQDs) was illustrated by using chemiluminescence, photoluminescence and ultraviolet analysis. Ranitidine can quench the chemiluminescence intensity of a luminol–H₂O₂–N,S‐CQDs system. So, a novel flow‐injection chemiluminescence method was designed to determine ranitidine within a linear range of 0.5–50 μg ml^?1 and a detection limit of 0.12 μg ml^?1. The method shows promising application prospects.