施氏鲟Asnanos1基因序列分析及其在雌雄不同组织中表达     

肖懿哲  朱友芳  孙玉龙  张鑫  张子平  王艺磊 《海洋科学》2017,41(4):17-23

本研究首先从施氏鲟(Acipenser schrenckii)性腺转录组中获得nanos1(Asnanos1)基因全长c DNA,并对其序列的准确性和特征分别进行PCR验证和生物信息学分析。进一步利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。利用荧光定量RT-PCR技术检测Asnanos1基因在雌、雄施氏鲟的性腺、心脏、鳃、脑、肾、肝脏、脾脏等组织中的表达量。结果显示:Asnanos1的c DNA全长为1 389 bp,其中,5′非编码区(UTR)为414 bp,3′UTR为279 bp,ORF为696 bp。该基因共编码231个氨基酸。Asnanos1基因在施氏鲟除心脏以外的各组织中均有表达,在雌、雄鱼的鳃、脑、肾、肝脏、脾脏等组织中表达量无显著差异,但在卵巢中的表达量显著高于其他各组织(P0.05)。所得到的结果可为人工培育全雌施氏鲟苗种提供一些基础资料。  相似文献   

Response of Planktonic and Benthic Microbial Community to Urban Pollution from Sewage Discharge in Jilin Reach of the Second Songhua River,China          下载免费PDF全文

Shanshan Lin  Ying Wang  Jifang Lin  Chengshi Quan 《洁净——土壤、空气、水》2014,42(10):1376-1383

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黄东海浮游生物群落呼吸率对碳平衡的重要性     

叶文琪  刘诚刚  蔡昱明  翟红昌  乐凤凤  王斌  寿鹿  陈全震  杜萍 《海洋与湖沼》2022,53(1):84-95

黄东海接受长江冲淡水和黑潮带来的大量营养盐和有机物质,其碳循环对陆架海碳源汇格局至关重要.浮游生物群落呼吸是影响碳循环的重要过程.为揭示黄东海浮游生物群落呼吸率(PCR)对碳平衡的贡献,于2011年四季使用黑白瓶培养法测定黄海南部及东海北部浮游群落呼吸率和初级生产力,并同步测定温度、盐度、营养盐、叶绿素和细菌丰度等环境...  相似文献   

翘嘴鲌(Culter alburnus)TMEM-57基因的克隆、组织表达和单核苷酸多态性分析     

祁鹏志  郭宝英  谢从新  吴常文  陆诗敏  段友健 《海洋与湖沼》2012,43(6):1196-1201

采用RT-PCR和快速扩增cDNA末端(rapid amplification of cDNA ends,RACE)技术首次克隆了翘嘴鲌TMEM-57蛋白基因的cDNA全序列,该序列全长为2822bp,其中5′-UTR区93bp,3′-UTR区731bp,开放阅读框(open reading frame,ORF)为1998bp,编码665个氨基酸。NJ法系统进化分析将同科或属中TMEM-57基因分成一个分支,表现出种属间的高度保守性。利用荧光定量PCR(qPCR)技术检测了不同组织的表达量,发现在卵巢中表达量最高,其次为心脏和脑。用直接测序法对TMEM-57基因部分区段单核苷酸多态性(SNP)进行了检测,寻找到11个新的SNP位点。  相似文献   

刺参虾青素基因的克隆及不同体色个体间表达差异的分析     

赵鹤凌  杨红生  赵欢  刘石林 《海洋科学》2012,36(3):22-28

仿刺参(Apostichopus japonicus Selenka)的体色变化很大,多数背腹均为黄褐色,极少数呈白化特征,背腹均为纯白色。经人工繁育实验发现,白化仿刺参的子代仍多具白化特征。本文研究了虾青素基因与刺参白化特征的相关性。在克隆虾青素基因cDNA全长的基础上,比较了普通仿刺参和白化仿刺参在不同发育时期虾青素基因表达量的差异。结果表明,该基因的cDNA含有2058个核苷酸,编码560个氨基酸。经实时定量PCR分析,白化成参体壁中虾青素基因表达量显著低于普通成参;而在仿刺参色素沉积的早期,白化稚参体壁中虾青素基因表达量从受精后第39天开始显著低于普通稚参。可见,仿刺参体壁中虾青素基因的低表达与刺参白化特征的发生密切相关。  相似文献   

Potential use of dissolved cyanobacterial DNA for monitoring toxic Microcystis cyanobacteria in filtered water     

《Physics and Chemistry of the Earth》2013

Toxic and non-toxic Microcystis sp. are morphologically indistinguishable cyanobacteria that are increasingly posing health problems in fresh water systems by producing odours and/or toxins. Toxic Microcystis sp. produces toxicologically stable water soluble toxic compounds called microcystins (MCs) that have been associated with cases of aquatic life and wildlife poisoning and kills including some cases of human illnesses/deaths around the world. Thus, the need for rapid detection of toxic Microcystis sp. in surface water is imperatively a necessity for early mitigation purposes. Genomic DNA from potentially toxic Microcystis sp. comprises of ten microcystin synthetase (mcy) genes of which six major ones are directly involved in MCs biosynthesis. In Polymerase Chain Reaction (PCR) methodsmcy genes can be amplified from intracellular/extracellular genomic DNA using PCR primers. However, little is known about the limitations of sourcing genomic DNA templates from extracellular DNA dissolved in water. In this work, filtered water (0.45 μM) from a Microcystis infested Dam (South Africa) was re-filtered on 0.22 μM syringe filters followed by genomic DNA isolation and purification from micro-filtrates (9 mL). Six major mcy genes (mcyABCDEG) from the isolated DNA were amplified using newly designed as well as existing primers identified from literature. PCR products were separated by gel electrophoresis and visualized after staining with ethidium bromide. The limitation of using dissolved DNA for amplification of mcy genes was qualitatively studied by establishing the relationship between input DNA concentrations (10.0–0.001 ng/μL) and the formation of respective PCR products. The amplification of mcyA gene using new primers with as little as 0.001 ng/μL of DNA was possible. Other mcy gene sensitivities reached 0.1 ng/μL DNA dilution limits. These results demonstrated that with appropriately optimized PCR conditions the method can provide accurate cost-effective tools for rapid detection of toxic Microcystis sp. in water giving early information for water quality monitoring against MC producing cyanobacteria.  相似文献   

Reliable Procedure for Molecular Analysis of Airborne Microflora in Three Indoor Environments: An Office and Two Different Museum Contexts     

Carole Gaüzère  Marina Moletta‐Denat  Faisl Bousta  Stéphane Moularat  Geneviève Orial  Sébastien Ritoux  Jean‐Jacques Godon  Enric Robine 《洁净——土壤、空气、水》2013,41(3):226-234

Biological aerosols from air constitute a significant source of exposure to microorganisms in public places. Airborne microorganisms are involved in the development of certain respiratory symptoms, allergies, or infections among users and occupants. Various sampling instruments have commonly been used in aerobiology to collect bacteria and fungi suspended in the air. The objective of this study was to develop a reliable procedure for sampling in indoor public environments presenting different levels of occupancy, airborne bacteria and fungi to be subjected to molecular analysis (bacteria and fungi quantitative PCR, capillary electrophoresis single strand conformation polymorphism fingerprinting). Four different sampling devices were tested in situ in an office building (open‐plan type) and the sampling strategy chosen was tested in two museum contexts. In accordance with the drawbacks involved to our study (quantitative and qualitative aspects, cost, and overcrowding), cyclone device appeared to be most suitable. The results underline the effectiveness of this high‐volume aerosol sampling device for both qualitative and quantitative molecular analysis. Four in situ sampling collections were carried out in 1 day in the Louvre Museum to study quantitative and qualitative variations of airborne bacterial and fungal diversity. The quantitative results revealed a similar order of magnitude for the numbers of both bacteria and fungi. In the Louvre Museum, the samples yielded between 3.7 × 10⁴ and 4.1 × 10⁴ genome equivalent (GE) bacteria/m³ air and between 5.0 × 10⁴ and 5.9 × 10⁴ GE fungi/m³ air and in the Decorative Arts Museum between, 2.1 × 10⁴ and 2.5 × 10⁴ GE bacteria/m³ air and between 1.4 × 10⁴ and 1.7 × 10⁴ GE fungi/m³ air. The results also indicate that the dominant bacterial community displayed a stable structure over a short period of time whereas dominant eukaryotic airborne community appeared more variable.  相似文献   

CYP1A mRNA expression in redeye mullets (Liza haematocheila) from Bohai Bay, China     

An L  Hu J  Yang M  Zheng B  Wei A  Shang J  Zhao X 《Marine pollution bulletin》2011,62(4):718-725

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Major human Hepatitis A virus genotype in Hong Kong marine waters and detection by real-time PCR     

Yang N  Chu DL  Wong MM  Qi H  Wu RS  Kong RY 《Marine pollution bulletin》2011,62(12):2654-2658

Marine waters from seven sites around Hong Kong with varying levels of sewage pollution were analyzed for Hepatitis A virus (HAV) by PCR cloning and DNA sequencing of the highly variable VP1/2A junction of the HAV genome. Phylogenetic analysis of 10 PCR clones from each of the HAV-positive marine sites indicated that human HAV genotype IB is the most widely distributed type in Hong Kong waters. A sensitive and quantitative TaqMan-based PCR method targeting the 5′-noncoding region (5′-NCR) of HAV was used to quantify HAV particles in marine water samples along with the total Escherichiacoli counts being enumerated on TBX medium for comparison. Our results showed that no correlation of any significance between HAV and E. coli counts was observed which underscores the inadequacy in using E. coli as a sanitary standard to predict the levels of HAV in marine waters.  相似文献   

大菱鲆亲子鉴定的微卫星多重PCR技术建立及应用     

苗贵东  杜民  杨景峰  徐营  樊廷俊  陈松林 《中国海洋大学学报(自然科学版)》2011,(Z1):97-106

用8个微卫星标记组合建立了2个微卫星多重PCR体系,对大菱鲆7个人工选育家系进行了系谱认证、亲子鉴定和遗传多样性研究。2个多重PCR体系中8个微卫星位点共检测到等位基因49个,每个位点等位基因数为3~8个。根据已知亲本及子代基因型,成功推断出了3个缺失亲本的基因型。在双亲未知的情况下2个多重PCR的8个微卫星位点累积排除概率是96.58%,已知1个亲本时累积排除率为99.71%,亲子鉴定准确率为96.42%。采用70个个体进行双盲验证,利用UPGMA法对7个家系的70个体进行了聚类分析,同一家系95.71%的个体聚类分析结果与系谱来源一致。Cervus 2.0软件亲子鉴定结果表明亲子鉴定准确率为92.86%。2个多重PCR体系的建立为大菱鲆不同家系混养后的亲子鉴定、系谱分析和分子辅助家系管理提供了技术手段。  相似文献