Biochemical and transcriptome analysis of small abalone Haliotis diversicolor to TBT exposure

由于海洋环境的巨大稀释能力，使得长期低浓度存在于海洋中的污染物所造成的生物生态效应很难在短时间内表现出来。但是，这种变化可在生物的细胞和分子水平上引起改变，如何识别这些变化，并通过这些变化分析可能造成的生物生态效应，是环境生物监测技术的研究重点。 本研究以杂色鲍（Haliotisdiversicolor）肝胰腺和血淋巴为材料，测定了0.35µg(Sn)/L三丁基锡（TBT）暴露2h，6h，24h，48h，96h和192h后的酸性和碱性磷酸酶（ACP和AKP），超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活力以及丙二醛（MDA）浓度，构建了鲍肝胰腺均一化cDNA文库，进行了大规模...Tributyltin (TBT) is one of the most disagreeable pollutants globally. Development of a biomarker for TBT detection is a pressing problem for environmental scientists. In this project, we investigated the effect of TBT exposure on the concentration of malondialdehyde (MDA) and the activity levels of the superoxide dismutase (SOD), catalase (CAT), and acid and alkaline phosphatase (ACP and AKP) ...学位：理学博士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：B20043401

Construction of Normalized cDNA Library from Hepatopancreas of Abalone Haliotis diversicolor supertexta Exposure to Tributyltin

用rdP试剂提取三丁基锡暴露诱导的杂色鲍肝胰腺总rnA,经OlIgOTEX纯化得到MrnA;应用SMArT技术合成双链CdnA,双链特异核酸酶(dSn)进行双链CdnA的均一化,构建了杂色鲍三丁基锡暴露诱导下的均一化CdnA文库。原始文库的库容为4.3x106Cfu/Ml,重组率为97.9%。从文库中随机挑选了3288个克隆进行测序,得到3048个高质量EST序列,其中有370条COnTIgS,2103条SInglETS,unIgEnES共2473条,冗余率为18.86%。以上结果说明该文库质量较好,为进一步筛选相关功能基因打下基础;较低的冗余率说明该文库值得继续使用大规模ESTS测序的方法寻找相关功能基因,并为进一步使用基因芯片技术研究相关功能基因的表达谱提供便利。A normalized cDNA library from hepatopancreas of abalone Haliotis diversicolor supertexta exposure to Tributyltin was constructed.Total RNA were prepared using RDP reagent.Oligotex(QIAGEN)was used to separate the mRNA from total RNA.The first strand cDNA was synthesized by transcription of mRNA with the SMART technique.The LD-PCR was performed using a modified SMART primer as the primer set,and the first-strand cDNA as the template to synthesize double strand cDNA.Double strand cDNA was normalized using Duplex-Specific Nuclease(DSN).The containing capacity of the unamplified cDNA library was 4.3×106 CFU/ml with a recombinant rate of 97.9%.A total of 3 288 clones were random selected to be sequenced,and 3 048 high quality ESTs were generated.After processing,a total of 2 473 unigenes comprising 370 contigs and 2 103 singlets were obtained.The redundancy was 18.86%.These results indicate that the normalized cDNA library is suitable to be used for further cloning and analysis of genes related to tributyltin exposure.厦门市科技计划项目(3502Z20055024);国家自然科学基金(20877034);集美大学创新团队基金(2008A001

Community structure of marine ultraphytoplankton in Xiamen waters using photosynthetic pigments

应用反相高效液相色谱(RP-HPLC)方法对厦门海域9个站位水样进行分析,对色素数据进行换算,结果表明在厦门海域超微型浮游植物优势类群是绿藻,而总浮游植物优势类群是硅藻;超微型浮游植物对总浮游植物生物量贡献为总生物量1.5%～11%,超微型粒级细胞在一些类群中(绿藻、金藻)中占有很大比重。在超微型浮游类群组成中没有甲藻纲,而蓝藻丰度也非常低。超微型浮游植物组成结构厦门港富营养化水域呈单优势类群结构,在九龙江河口区域受冲淡水与陆源成分影响,结构较为复杂。Photosynthetic pigments were measured by RP-HPLC in seawaters samples of Xiamen in winter(December, 2002), and the diagnostic pigment data were converted into chlorophyll a biomass to assess the community structure both in total and ultra-size phytoplankton. The result showed that chlorophyte was dominant group in ultra-phytoplankton community, while diatoms was prevaided in total phytoplankton community. The biomass of ultra-phytoplankton only occupied about 1.5% to 11% biomass of all phytoplankton(indicated by Chlorophyll a), however, the ultra-size phytoplankton contributed a large proportion of biomass in chlorophyte and prymnesiaphyceae. Both dinophyceae and cyanobacteria were not detected in ultraphytoplankton community. Community structure of ultraphytoplankton was simple in eutrophic aquicultral waters, while complicated in Jiulong River estuary.国家自然科学基金(40276039,40076031

cDNA Library Construction of Testis and Ovary ofScylla serrata with SMART Technique

【中文摘要】 采用RDP试剂(改进自ChomczynskiandSacchi1987)提取锯缘青蟹精巢和卵巢的总RNA,经oligotex试剂盒纯化得到mRNA.利用SMART(switchingmechanismat5'endofRNAtranscript)技术,构建成精巢和卵巢的全长cDNA文库.在构建好的文库中,精巢的文库含有约1.45×106的重组子,卵巢的文库含有约为1.75×106的重组子.插入cDNA长度为500bp～3kb,说明两文库均具有良好的质量,为进一步筛选、克隆精巢与卵巢特异表达基因奠定了基础. 【英文摘要】 To clone the testis and ovary specific expression genes of Scylla serrata,we constructed two full-length cDNA libraries using the SMART cDNA library construction kit(Clontech).Total RNA were prepared from testes and ovaries using RDP,a reagent modified from Chomczynski and Sacchi(1987)for the simultaneous isolation of RNA,DNA,and protein.Oligotex(QIAGEN)was used to separate the mRNA of testes and ovaries from total RNA.The"anchor first-strand cDNA"containing asymmetricalSfiⅠrestriction enzyme sites(A&B) was...国家教育部骨干教师基金;福建省重中之重项目资

cDNA Library Construction of Testis and Ovary ofScylla serrata with SMART Technique

采用RDP试剂(改进自ChomczynskiandSacchi1987)提取锯缘青蟹精巢和卵巢的总RNA,经oligotex试剂盒纯化得到mRNA.利用SMART(switchingmechanismat5'endofRNAtranscript)技术,构建成精巢和卵巢的全长cDNA文库.在构建好的文库中,精巢的文库含有约1.45×106的重组子,卵巢的文库含有约为1.75×106的重组子.插入cDNA长度为500bp～3kb,说明两文库均具有良好的质量,为进一步筛选、克隆精巢与卵巢特异表达基因奠定了基础. 【英文摘要】 To clone the testis and ovary specific expression genes of Scylla serrata,we constructed two full-length cDNA libraries using the SMART cDNA library construction kit(Clontech).Total RNA were prepared from testes and ovaries using RDP,a reagent modified from Chomczynski and Sacchi(1987)for the simultaneous isolation of RNA,DNA,and protein.Oligotex(QIAGEN)was used to separate the mRNA of testes and ovaries from total RNA.The"anchor first-strand cDNA"containing asymmetricalSfiⅠrestriction enzyme sites(A&B) was...国家教育部骨干教师基金;; 福建省重中之重项目资

JUNO Sensitivity on Proton Decay p→νˉK+p\to \bar\nu K^+ Searches

The Jiangmen Underground Neutrino Observatory (JUNO) is a large liquid scintillator detector designed to explore many topics in fundamental physics. In this paper, the potential on searching for proton decay in $p\to \bar\nu K^+$ mode with JUNO is investigated.The kaon and its decay particles feature a clear three-fold coincidence signature that results in a high efficiency for identification. Moreover, the excellent energy resolution of JUNO permits to suppress the sizable background caused by other delayed signals. Based on these advantages, the detection efficiency for the proton decay via $p\to \bar\nu K^+$ is 36.9% with a background level of 0.2 events after 10 years of data taking. The estimated sensitivity based on 200 kton-years exposure is $9.6 \times 10^{33}$ years, competitive with the current best limits on the proton lifetime in this channel

JUNO sensitivity on proton decay p → ν K + searches*

The Jiangmen Underground Neutrino Observatory (JUNO) is a large liquid scintillator detector designed to explore many topics in fundamental physics. In this study, the potential of searching for proton decay in the $p\to \bar{\nu} K^+$ mode with JUNO is investigated. The kaon and its decay particles feature a clear three-fold coincidence signature that results in a high efficiency for identification. Moreover, the excellent energy resolution of JUNO permits suppression of the sizable background caused by other delayed signals. Based on these advantages, the detection efficiency for the proton decay via $p\to \bar{\nu} K^+$ is 36.9% ± 4.9% with a background level of $0.2\pm 0.05({\rm syst})\pm$$0.2({\rm stat})$ events after 10 years of data collection. The estimated sensitivity based on 200 kton-years of exposure is $9.6 \times 10^{33}$ years, which is competitive with the current best limits on the proton lifetime in this channel and complements the use of different detection technologies