红砂正常和脱水组织中总RNA提取的改进CTAB法   总被引：11，自引：5，他引：6  

刘玉冰  张腾国  安黎哲  王刚 《中国沙漠》2006,26(4):600-603

红砂属于柽柳科超旱生小灌木,是荒漠半荒漠地区植被中的优势种。为了研究促分裂原激活的蛋白激酶(MAPK)信号传导通路在这类植物抗旱中的作用,我们尝试了很多RNA提取的方法都没有获得完整的RNA。在脱水组织中由于多糖、多酚以及其他与核酸能结合或共沉淀的化合物的含量升高而更不容易得到高质量的RNA。以CTAB为基础的改进的RNA提取方法,能成功地从少量的红砂植物的正常组织和脱水组织中获得高质量的RNA。这种方法经济且操作方便,提取的RNA纯度和完整性都很好,可用于RT-PCR反应并成功克隆了550 bp 左右的MAPK基因片段。  相似文献   

单条固定线虫基因组DNA提取及18S rRNA基因PCR扩增   总被引：2，自引：0，他引：2  

沈锡权  杨官品  廖梅杰 《海洋科学》2005,29(5):33-36

根据线虫18S核糖体RM基因PCB扩增效果比较了丙酮、乙醇、乙醇 0．05mol／L FDTA(pH8．0)和5％海水福尔马林4种固定剂,碱裂解和蛋白酶K处理2种单条线虫基因组DNA提取方法的优劣。用乙醇固定的样品最适合制备RR模板DNA,而5％海水福尔马林固定的样品能最完整地保持样品形态。蛋白酶K处理获得的DNA较碱裂解获得的更适合PCR扩增。结果有助于分子生物学方法在海洋线虫分类、多样性和生态学研究中的应用。  相似文献   

Analysis of Effects of Berberine on the Photosynthesis of Microcystis aeruginosa at Gene Transcriptional Level          下载免费PDF全文

Xiangdong Bi  Shulin Zhang  Wei Dai  Kezhi Xing  Yue Pang  Dajuan Zhang 《洁净——土壤、空气、水》2015,43(1):44-50

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Cloning,characterization, and expression of Cytochrome b (<Emphasis Type="Italic">Cytb</Emphasis>)—a key mitochondrial gene from <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis>     

赵丽媛  米铁柱  甄毓  于志刚 《中国海洋湖沼学报》2012,30(3):424-432

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一种基于干扰肌动蛋白基因导致方形网纹溞(Ceriodaphnia quadrangula)死亡的RNA干扰(RNAi)模型构建     

陈宏健  谢松  李理想  柳峰松 《海洋与湖沼》2014,45(2):322-327

方形网纹溞(Ceriodaphnia quadrangula)可作为研究水生动物生物学的良好模式动物。为探索细菌介导表达的双链RNA(dsRNA)对方形网纹溞进行RNA干扰(RNAi)技术是否可行,本研究构建了方形网纹溞肌动蛋白(actin)基因的干扰载体Actin-L4440,并将其转入大肠杆菌HT115。此宿主菌经异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导后能表达actin基因部分片段的dsRNA。将此宿主菌投喂方形网纹溞,能抑制其体内actin基因的表达,并导致方形网纹溞死亡率明显提高。本研究成功构建了方形网纹溞的RNAi模型,为以后的研究提供了一定的参考。  相似文献   

慢性氨氮胁迫对鲻鱼(Mugil cephalus)幼鱼组织细胞免疫指标的影响研究   总被引：1，自引：0，他引：1  

蒋玫  李磊  沈新强  吴庆元  牛俊翔 《海洋与湖沼》2014,45(3):529-535

以鲻鱼(Mugil cephalus)幼鱼为研究对象,研究低浓度氨氮长时间胁迫对组织细胞免疫指标和遗传物质代谢的影响。实验分0.35、0.7、1.5和3mg/L氨氮处理组,分别于0、5、10、15、20d取样并进行相关指标测定。结果显示:肝脏和鳃丝的丙二醛(MDA)含量表现出先上升后下降的趋势,MDA活性与氨氮浓度呈一定的正相关。氨氮对MDA的影响具有不同组织和不同时序的选择性。不同浓度氨氮胁迫下,鳃丝和肝脏Na+-K+-ATP活性均呈现先升高后降低,并随着时间的增加逐渐趋于稳定。Na+-K+-ATP活性与氨氮浓度呈一定的负相关性,尤其是3mg/L处理组与对照组差异性显著(P0.05)。在氨氮浓度0.7mg/L和1.5mg/L时,肌肉中RNA/DNA均表现出不同程度的增加,各浓度组与对照组差异不显著;而高浓度(3mg/L)比值基本无变化。肝脏和鳃丝的Na+-K+-ATP酶的mRNA表达量,在氨氮胁迫第5d时检测到达最高,随后表达量缓慢回落,实验20d除3mg/L处理组表达量仍高于对照组水平外,其他处理组均降至对照组水平。Na+-K+-ATP酶的表达水平变化趋势与Na+-K+-ATP活性基本一致。结果表明,在一定浓度的氨氮长时间胁迫作用下,抗氧化免疫系统和肌肉中的核酸代谢受到了一定的影响。  相似文献   

海洋无脊椎动物组织总RNA提取方法的探讨   总被引：2，自引：0，他引：2  

冯政夫  王琳  李文侠  部凡  胡彦江  朱伟 《海洋科学》2014,38(11):24-28

RNA提取是分子生物学研究的基础实验,由于海洋无脊椎动物组织结构以及体成分的特殊性,导致现有的RNA提取方法不能完全胜任此工作。作者以刺参(Apostichopus japonicus)、栉孔扇贝(Chlamys farreri)和凡纳滨对虾(Litopenaeus vannamei)为研究对象,对异硫氰酸胍RNA提取方法进行了改进。栉孔扇贝和凡纳滨对虾的肝胰腺富含RNase,刺参体壁结构致密、富含胶原蛋白,而成熟期精巢富含多糖,因此在这些动物组织中提取RNA时,在原有方法基础上增加了瞬时匀浆、增加氯仿及酸性酚的抽提次数,或添加高盐溶液等方法。实验结果显示,改进方法后获得的RNA具有完整的28S、18S和5S r RNA条带,A260nm/A280nm和A260nm/A230nm比值分别为1.89~2.0和1.98~2.11,β-actin基因扩增条带清晰,所获得的组织RNA的纯度和质量符合c DNA合成和基因功能研究的要求。  相似文献   

Spatial distribution, diversity and composition of bacterial communities in sub-seafloor fluids at a deep-sea hydrothermal field of the Suiyo Seamount     

Shingo Kato  Kurt Hara  Hiroko Kasai  Takashi Teramura  Michinari Sunamura  Jun-ichiro Ishibashi  Takeshi Kakegawa  Toshiro Yamanaka  Hiroyuki Kimura  Katsumi Marumo  Tetsuro Urabe  Akihiko Yamagishi   《Deep Sea Research Part I: Oceanographic Research Papers》2009,56(10):1844-1855

Spatial distribution, diversity, and composition of bacterial communities within the shallow sub-seafloor at the deep-sea hydrothermal field of the Suiyo Seamount, Izu-Bonin Arc, Western Pacific Ocean, were investigated. Fluids were sampled from four boreholes in this area. Each borehole was located near or away from active vents, the distance ranging 2–40 m from active vents. In addition, fluids discharging from a natural vent and ambient seawater were sampled in this area. We extracted DNA from each sample, amplified bacterial 16S rRNA genes by PCR, cloned the PCR products and sequenced. The total number of clones analyzed was 348. Most of the detected phylotypes were affiliated with the phylum Proteobacteria, of which the detection frequency in each clone library ranged from 84.6% to 100%. The bacterial community diversity and composition were different between hydrothermal fluids and seawater, between fluids from the boreholes and the vent, and even among fluids from each borehole. The relative abundances of the phylotypes related to Thiomicrospira, Methylobacterium and Sphingomonas were significantly different among fluids from each borehole. The phylotypes related to Thiomicrospira and Alcanivorax were detected in all of the boreholes and vent samples. Our findings provide insights into bacterial communities in the shallow sub-seafloor environments at active deep-sea hydrothermal vent fields.  相似文献   

Changes in the Chemical Components of Light Crude Oil During Simulated Short Term Weathering     

MA Qimin )  LUO Xia)  and YU Zhigang) ) College of Environmental Science  Engineering  Ocean University of China  Qingdao  P. R. China ) College of Chemistry  Chemical Engineering  Ocean University of China  Qingdao  P. R. China 《中国海洋大学学报(英文版)》2007,6(3):231-236

To unambiguously identify spilled oils and to link them to the known sources are extremely important in settling ques-tions of environmental impact and legal liability. The fate and behavior of spilled oils in the environment depend on a number of physicochemical and biological factors. This paper presents the results regarding changes in chemical composition of light crude oil during simulated short-term weathering based on natural environmental conditions. The results show that the saturated hydrocarbons of the light crude oil mainly distribute between n-C8 and n-C23 and the most abundant n-alkanes are found in the n-C10 to n-C16. The main chemical components of the light crude oil are n-alkanes and isoprenoids. The aromatic compounds are subordinate chemical components. Under the conditions of the weathering simulation experiment,n-alkanes less than n-C12,toluene and 1,3-dimethyl ben-zene are lost after 1 d weathering,the n-C13,n-C14,naphthalene and 2-methyl-naphthalene are lost on the fifth day of weathering,and n-C15 alkane components show certain weatherproof capability. The ratios n-C17/pristane and n-C18/phytane are unaltered and can be used to identify the source of the light crude oil during the first 8 d of weathering. After 21 d,the ratio pristine/phytane can not pro-vide much information on the source of the spilled light crude oil. Triterpanes(m/z 191) as biomarker compounds of light crude oil are more valuable.  相似文献   

Temperature Influences on the Expression of GFP Promoted by the Upstream Sequence of cpcB from Arthrospiraplatensis     

LU Yongzhong ZHANG Xuecheng 《中国海洋大学学报(英文版)》2007,6(3):244-248

In order to investigate the regulation mechanism of the phycocyanin gene,a series of functional analyses of the upstream sequence of cpcB gene from Arthrospira platensis were conducted in E. coli with green fluorescent protein encoding gene(gfp) as the reporter. Results showed that the gfp gene could express at a high level under the promotion of the upstream sequence,suggesting the existence of some strong promoter elements in it. The expression of GFP was influenced by temperature. Higher temperature led to higher expression level. The bioinformatics analyses followed by mutation analyses on the secondary structure of translation ini-tiation region(TIR) revealed that RNA thermosensor might account for the temperature regulation.  相似文献