大黄鱼(Larimichthys crocea)白细胞介素10(IL-10)基因克隆及溶藻弧菌(Vibrio alginolyticus)侵染后表达变化分析

白细胞介素10(interleukin10,IL-10)是鱼类先天性免疫因子之一,在炎症反应和免疫调节中有着重要作用。本研究获得大黄鱼(Larimichthys crocea)IL-10基因cDNA序列,由899个核苷酸组成,含1个555bp的开放阅读框、编码184个氨基酸,预测编码蛋白分子量为21.24kDa、N端有22个氨基酸组成的信号肽序列。氨基酸序列多重比对表明大黄鱼IL-10具有IL-10家族特征性序列和构成2对二硫键的4个保守性半胱氨酸,与欧洲鲈(Dicentrarchuslabrax)IL-10序列相似性最高(78.5%);系统进化树分析显示大黄鱼IL-10和其他鱼类IL-10形成一个大簇,与欧洲鲈进化关系最近。荧光定量PCR(quantitative real-time PCR, qRT-PCR)检测结果显示健康大黄鱼IL-10基因在鳃和肠中高表达,肝和头肾中低表达;溶藻弧菌(Vibrio alginolyticus)侵染后大黄鱼肠、脑、脾、肝和头肾中IL-10基因表达量均显著上调(P0.05),其中头肾和肝中上调幅度最大(分别为菌侵染前的10.06倍和8.79倍)。综上,大黄鱼IL-10基因表达与病原菌感染密切相关,为深入研究大黄鱼IL-10的生物学功能及免疫机制提供了基础资料。     

梅氏新贝尼登虫(Neobenedenia melleni)卵黄铁蛋白的cDNA克隆、原核表达及抗血清制备

从cDNA文库中获得了梅氏新贝尼登虫卵黄铁蛋白(NmYF)的编码序列。序列全长739个核苷酸,3′-末端具有polyA尾,单一大开放阅读框编码一个由226个氨基酸组成的分子量为26.1kDa的蛋白。序列比较表明,NmYF与卫氏并殖吸虫卵黄铁蛋白最相似,氨基酸序列同源性为23.7%。系统进化树分析表明,NmYF、卫氏并殖吸虫卵黄铁蛋白和头槽绦虫卵黄铁蛋白形成了一个小的进化簇,而体铁蛋白序列形成了一个大的进化簇,揭示了卵黄铁蛋白和体铁蛋白间的差异以及进化上的相关性。这是梅氏新贝尼登虫卵黄铁蛋白基因序列的首次报道。随后,构建了插入有全长NmYF基因开放阅读框序列的原核表达质粒pET-22b-NmYF,转化大肠杆菌BL21pLysE菌株,经IPTG诱导表达后,SDS-PAGE检测表明预期大小的目的蛋白大量表达。随后制备了目的蛋白的多克隆抗血清,它能与目的蛋白起强的特异性反应,但与细菌自身蛋白不起反应,可用于后续研究。     

日本鬼鲉(Inimicus japonicus)线粒体Cyt b和COⅡ基因的克隆及鲉形目系统发育分析

根据鲉形目鱼类Cyt b和CO Ⅱ上下游保守序列各自设计引物,对日本鬼鲇的线粒体DNA进行PCR扩增,克隆并测定了Cyt b和COⅡ因的全序列,CO Ⅱ基因为691bp,Cyt b为1141bp.从GenBank下载28种鲉形目及外群鱼类的Cyt b部分或全序列和10种形目及外群鱼类的COⅡ部分或全序列,然后进行比对分...     

中国暖季短时强降水分布和日变化特征及其与中尺度对流系统日变化关系分析

短时强降水是强对流天气的一类.基于中国国家气象信息中心质量控制后的1991-2009年876个基本基准气象站整点逐时降水资料,通过不同时段的发生时次频率分析,给出了中国暖季(4-9月)不小于10、20、30、40、50 mm/h短时强降水的时空分布特征,并重点同利用静止气象卫星红外相当黑体亮度温度(T_BB)资料获得的中尺度对流系统(MCS)日变化特征进行了对比分析.结果表明,中国短时强降水时次频率地理分布同暴雨(≥50mm/d)分布都非常相似,但50mm/h以上的短时强降水时次频率非常低,地理分布差异显着.短时强降水发生频率最高的区域为华南,其次为云南南部、四川盆地、贵州南部、江西和长江下游等地.最大降水强度可超过180mm/h(海南);在短时强降水发生频率很低的区域,也有超过50mm/h的强降水.从月际变化来看,7月最为活跃,其次为8月.逐候变化显示,短时强降水具有显着的间歇性发展特征(跳跃性分布的特征),但总体上呈现缓慢增强、迅速减弱的特点;以7月第4候最为活跃.中国总体平均的短时强降水的频率和最大强度的日变化有3个峰值,主峰在午后(16-17时,北京时),次峰在午夜后(01-02时)和早晨(07-08时);中午前后(10-13时)最不活跃.中国短时强降水和中尺度对流系统的日变化特征基本一致,但午夜后时段二者存在较大差异.不同区域的短时强降水和中尺度对流系统日变化具有不同的活跃时段和传播特征,具有单峰型、双峰型、多峰型和持续活跃型等日变化类型,这不仅与较大尺度的天气系统环流相关,且与地势、海陆等地理分布密切相关.     

A CLIMATOLOGY OF DEEP CONVECTION OVER SOUTH CHINA AND THE ADJACENT WATERS DURING SUMMER

Due to the higher temporal and spatial resolution and the better integrality of long-term satellite infrared(IR) Brightness Temperature(TBB) data,a climatology of deep convection during summer over South China and the adjacent waters is presented in this paper based on the 1-hourly infrared IR TBB data during June-August of 1996-2007(except 2004).The results show that the geographic distribution of deep convection denoted by TBB ≤-52℃ over South China and the adjacent waters are basically consistent with previous statistical results based on surface thunderstorm observations and low-orbit satellite lightning observations.The monthly,ten-day,five-day and diurnal variations of deep convection in this region are focused on in this paper.There are 5 active deep-convection areas in June-August.The monthly variations of the deep convection are closely associated with the large-scale atmospheric circulations.The deep convection over the land areas of South China is more active in June while that over the South China Sea is more active in July and August.The development of deep convection is prominently intermittent and its period is about 3 to 5 five-day periods.However,the deep convection over the coastal areas in South China remains more active during summer and has no apparent intermittence.The ten-day and five-day variations of deep convection show that there are different variations of deep convection over different areas in South China and the adjacent waters.The tendency of deep convection over the land areas of South China is negatively correlated with that over the South China Sea.The diurnal variations of deep convection show that the sea-land breeze,caused by the thermal differences between land and sea,and the mountain-valley breeze,caused by the thermal differences between mountains and plains or basins,cause deep convection to propagate from sea to land in the afternoon and from land to sea after midnight,and the convection over mountains propagates from mountains to plains after midnight.The different diurnal variations of deep convection over different underlying surfaces show that not only there are general mountainous,marine and multi-peak deep convection,but also there is longer-duration deep convection over coastal areas and other deep convection triggered and maintained by larger-scale weather systems in South China during summer.     

PaCLR介导PaLECT2激活脂多糖刺激的香鱼(Plecoglossus altivelis)单核巨噬细胞功能

香鱼(Plecoglossus altivelis)白细胞衍生趋化因子2(leukocyte cell-derived chemotaxin 2,PaLECT2)可以激活香鱼头肾来源的单核巨噬细胞(monocytes/macrophages,MO/MΦ),提高细菌感染后香鱼存活率。进一步研究显示,PaLECT2与香鱼C型凝集素受体(C-type lectin receptor,PaCLR)相互作用可激活静息状态下香鱼MO/MΦ,增强其免疫活性。在前期基础上,本研究通过抗体封闭实验检测PaCLR是否介导PaLECT2激活LPS刺激的MO/MΦ功能。结果显示,重组PaLECT2成熟肽(rPaLECT2m)显著提高了LPS刺激的香鱼MO/MΦ细胞因子的表达,并增强了其吞噬能力和杀菌活性。封闭PaCLR显著抑制了rPaLECT2m激活LPS刺激的香鱼MO/MΦ细胞因子表达、吞噬和杀菌功能。因此,PaCLR介导PaLECT2可以激活病理状态香鱼MO/MΦ。这一认识进一步揭示了LECT2在病理状态的鱼类巨噬细胞调控和炎症反应中的作用及机制。     

香鱼(Plecoglossus altivelis)铁蛋白的分子鉴定、表达及功能研究

铁蛋白(Ferritin,Fer)是一类广泛存在于生物体内、并与铁代谢密切相关的铁储存蛋白。本研究通过香鱼(Plecoglossus altivelis)单核/巨噬细胞转录组测序获得铁蛋白亚基基因(Pa Fer)c DNA全序列,结果显示,Pa Fer由943个核苷酸组成,开放阅读框为531bp,编码176个氨基酸,预测相对分子质量为20.4k Da,等电点为5.46。氨基酸序列多重比对结果显示,Pa Fer具有保守的铁蛋白样双铁结构域,与虹鳟(Oncorhynchus mykiss)和大西洋鲑(Salmo salar)Fer M亚基(Fer-M)同源性最高,均为92%;系统进化树分析表明,Pa Fer与胡瓜鱼、大西洋鲑和虹鳟的Fer-M聚为一个小簇,与虹鳟Fer-M进化相关性最高。实时荧光定量PCR(quantitative real-time PCR,q PCR)结果表明,Pa Fer m RNA主要在香鱼脾脏中表达量最高,肾脏、鳃、脑和心脏中次之;鳗弧菌(Vibrio anguillarum)感染后,香鱼肝脏、脾脏和肾脏等组织中Pa Fer m RNA表达显著上调。原核表达了Pa Fer重组蛋白(r Pa Fer),r Pa Fer以包涵体形式表达,镍柱亲和层析纯化重组蛋白,尿素梯度透析法复性r Pa Fer。铁螯合实验结果显示,当r Pa Fer浓度为4μg/m L时,螯合铁的能力最大,为43.96%。抑菌实验结果表明,r Pa Fer对鳗弧菌、溶藻弧菌(Vibrio alginolyticus)和创伤弧菌(Vibrio vulnificus)的最小抑菌浓度分别为1.5625、25.0和100.0μg/m L。综上,Pa Fer与香鱼抗病原感染的免疫反应紧密相关,可能通过调控铁代谢参与鱼类抵抗微生物的宿主自身免疫过程。本研究为进一步研究鱼类Fer的功能及在病原菌感染免疫中的分子机制奠定基础。     

CMA-GFS 4DVar边界层过程线性化的改进北大核心CSCD

持续发展和优化切线性模式的线性化物理过程,保持与非线性模式一致是改善四维变分同化(4DVar)分析和预报效果的有效方法之一。目前业务系统的CMA-GFS模式采用基于Charney-Phillips(C-P)跳点的边界层参数化方案,而CMA-GFS 4DVar系统中采用基于Lorenz跳点的边界层线性化方案。为改善CMA-GFS 4DVar系统的边界层分析和预报效果,基于C-P跳点的边界层参数化方案研发了新边界层线性化方案,并通过对方案中地表热量通量和水汽通量扰动、自由大气的理查逊系数扰动、边界层的热量和动量交换系数扰动等进行更加精细地规约化约束,在确保CMA-GFS切线性和伴随模式稳定运行的情况下,减少线性化过程对切线性模式预报精度的影响。切线性近似试验检验表明:相较于原方案,新边界层线性化方案可以减少边界层位温和比湿的相对误差,最大可减少10%。批量4DVar循环同化试验表明:新边界层线性化方案可以有效改善切线性模式对低层位温、风场和比湿扰动的预报精度,减少4DVar内外循环目标泛函的相对差异,并提高700 hPa位势高度的可预报时效。     

Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters

Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection (LOD) of 0.5 ng of genomic DNA (orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples.     

海水养殖环境中拟除虫菊酯类农药降解菌的分离鉴定及降解特性研究

从近岸海水养殖环境中分离获得一株能高效降解氯氰菊酯(Cypermethrin,CYP)和溴氰菊酯(Deltamethrin,DEL)的菌株HS-10,经生理生化和16S rDNA分析,初步鉴定其为假交替单胞菌(Pseudoalteromonas sp.)。不同降解条件下的实验结果表明,菌株HS-10在pH 7.0、温度28°C的环境中具有较好的生长和降解效率,在该条件下,对初始浓度为100mg/L的CYP和DEL的降解效率分别为75.6%和90.9%。进一步采用发光细菌和核磁共振(NMR)方法对菌株HS-10的降解产物进行分析,结果表明,CYP和DEL主要通过微生物降解消除,其降解产物的毒性显著降低。同时,考察了菌株HS-10对氯氰菊酯、溴氰菊酯、联苯菊酯(BIF)、氟氯氰菊酯(CYF)和氰戊菊酯(FEN)5种拟除虫菊酯类农药(浓度分别为100mg/L)的降解效率,结果表明,对5种农药的降解率均达到50%以上,本研究获得的菌株HS-10可用于海水养殖环境中拟除虫菊酯类农药残留污染的生物修复。