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本文以长牡蛎27个幼虫发育阶段个体以及成体的5个组织织作为实验材料,采用实时荧光定量PCR技术对Dmrt家族中的2个基因（CgDsx和CgDmrtA2）的主要对长牡蛎Dmrt家族中的2个基因（CgDsx和CgDmrtA2）在幼虫时期和成体组织中的表达模式以及在性别决定中可能发挥的作用进行了研究。采用实时荧光定量PCR技术对幼虫27个发育阶段以及成体5个组织的表达进行了测定。结果表明,长牡蛎CgDsx基因在胚胎发育初期有大量表达,其中囊胚期到担轮幼虫初期表达量最高从囊胚期到担轮幼虫初期表达量最高,其后之后表达量开始降低,在D形幼虫后期之后一直维持在极低的表达水平,此后仅在成体的雄性性腺中具有高度表达。该结果表明由此可见,CgDsx 可能也对早期胚胎发育起一定调控作用,除了同时参与了性别决定外,可能也对早期胚胎发育起一定调控作用。CgDmrtA2在长牡蛎所有组织中均有表达,各组织间表达差异不显著,在长牡蛎D形幼虫至壳顶后期表达量较高,表明其参与了胚胎中后期的发育过程说明它参与了胚胎中后期的发育过程,可能与神经的形成相关,但是其具体功能还需要进一步研究。 相似文献
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对虾白斑综合症病毒（white spot syndrome virus,WSSV）是危害对虾养殖业的主要病原,而WSSV极早期基因对病毒功能基因的转录翻译和调控起着至关重要的作用.本研究将WSSV基因组以超声法随机断裂至400～900bp DNA片段,并插入启动子缺失且多克隆位点下游具有报告基因的载体来构建文库,转染昆虫细胞Hi5.倒置荧光显微镜观察发现部分转染后的细胞有增强型绿色荧光蛋白（EGFP）的表达,表明这部分细胞中的DNA片段可能具有极早期启动子活性.流式细胞仪分析显示,这部分荧光细胞占总细胞的比例为0.35%.由于转染效率为50%,可认为占比0.7%的WSSV基因组随机片段能在Hi5细胞中激发下游报告基因的转录翻译,可能具有极早期基因启动子活性.这对于进一步筛选和鉴定WSSV极早期基因及开展极早期启动子研究具有重要的意义. 相似文献
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为发掘罗氏沼虾(Macrobrachiumrosenbergii)卵巢发育过程中的重要功能基因,采用IlluminaHiSeqTM4000高通量测序平台对罗氏沼虾卵巢发育四个时期卵巢组织进行转录组测序。所测序列经质控、组装后比对到NR、String、Swiss-prot、Pfam、GO、KEGG数据库中注释,并进行差异基因聚类分析。结果显示,四个样品共产生221580604个Cleanreads,拼接获得95379个Unigenes。分别对四个样品测序文库进行两两比较,共检测到6605个差异表达基因,其中显著上调基因2410个,显著下调基因4195个。GO功能分类显示,共有6422条unigenes可分为分子功能、细胞组分、生物学过程3大类59分支,差异基因主要涉及糖类转运、氨基酸合成、酶活性、细胞膜组成等。通过KEGG通路注释,共有8423条unigenes被注释,涉及184种代谢途径,有7个代谢通路与卵巢发育相关,差异基因的KEGG富集结果显示,药物代谢细胞色素P450通路、氨基丁酸神经元突触、鞘糖脂生物合成、氮素代谢等通路富集显著。此外,对SSR与SNP分子标记进行了鉴定。研究结果为进一步探索罗氏沼虾卵巢发育机制提供了基础信息。 相似文献
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Saccharina japonica is a common macroalga in sublittoral communities of cold seawater environments,and consequently may have highly effi cient ribulose-1,5-bisphosphate carboxylase/oxygenase(Rubisco)activity for carbon assimilation.In our study,we cloned the full-length Rubisco gene from S.japonica(SJ-rbc).It contained an open reading frame for a large subunit gene(SJ-rbcL)of 1 467 bp,a small subunit gene(SJ-rbcS)of 420 bp,and a SJ-rbcL /S intergenic spacer of 269 bp.The deduced peptides of SJ-rbcL and SJ-rbcS were 488 and 139 amino acids with theoretical molecular weights and isoelectric points of 53.97 kDa,5.81 and 15.84 kDa,4.71,respectively.After induction with 1 mmol/L isopropyl-β-Dthiogalactopyranoside for 5 h and purifi cation by Ni 2+ affi nity chromatography,electrophoresis and western blot detection demonstrated successful expression of the 55 kDa SJ-rbcL protein.Real-time quantitative PCR showed that the mRNA levels of SJ-rbcL in gametophytes increased when transferred into normal growth conditions and exhibited diurnal variations: increased expression during the day but suppressed expression at night.This observation implied that Rubisco played a role in normal gametophytic growth and development.In juvenile sporophytes,mRNA levels of SJ-rbcL,carbonic anhydrase,Calvin-BensonBassham cycle-related enzyme,and chloroplast light-harvesting protein were remarkably increased under continuous light irradiance.Similarly,expression of these genes was up-regulated under blue light irradiance at 350 μmol/(m 2·s).Our results indicate that long-term white light and short-term blue light irradiance enhances juvenile sporophytic growth by synergistic effects of various photosynthetic elements. 相似文献
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三（2-氯乙基）磷酸酯（Tris(2-carboxyethyl)phosphine, TCEP）是一种新型内分泌干扰物（Endocrine Disrupting Chemicals,EDCs),低剂量就会造成很强的生物毒性。通过研究不同质量浓度梯度的TCEP对养殖大菱鲆的胁迫,发现其在24 h 、48 h 、72 h 和96 h的半致死浓度（LC50）分别为190.76 mg/L、159.94 mg/L、140.70 mg/L和110.71 mg/L;安全浓度（SC）为33.60 mg/L。24 h内不同质量浓度梯度的TCEP和不同胁迫时间对养殖大菱鲆的影响表明,高于质量浓度33.60 mg/L的TCEP对大菱鲆具有急性毒性效应。用实时荧光定量PCR技术分析TCEP急性胁迫的养殖大菱鲆体内2种细胞免疫因子-组织相容性复合体alpha（MHC-Ⅱ α）和肿瘤坏死因子alpha（TNF-α）的基因相对表达水平,发现经TCEP胁迫后2种细胞免疫因子的表达量都显著上调;随TCEP的质量浓度升高,特别是达到47.69 mg/L（$ \frac{1}{4} $ LC50）以上时表达量的变化更加明显;而胁迫时间超过12 h,基因表达量会呈现先上升后下降的趋势,说明对大菱鲆免疫系统造成急性损伤。研究结果可为大菱鲆健康养殖提供参考,并为环境污染物检测提供分子标记参考。 相似文献
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祁连山高寒草原碱性土壤固氮微生物数量及固氮基因(nifH)群落结构研究 总被引：2，自引：1，他引：1
对祁连山高寒草原碱性土壤可培养固氮菌数量、 固氮基因(nifH)群落结构及其理化性质进行了研究. 结果表明: 固氮菌数量在土层0~40 cm处于3.6×105~0.21×105CFUs·g-1之间, 除了海拔3 001 m的土样AQ4外, 其他3个土样固氮菌数量随着土壤深度的加深而减小. 固氮菌数量与地下生物量呈显著正相关性, 与有机碳、 可溶有机氮、 速效磷和速效钾呈正相关性, 而与pH值和全盐呈负相关性. 通过基因测序得到的37个固氮基因nifH克隆中, 蓝藻门占41%, 变形菌门占8%, 厚壁菌门占14%, 未知菌株占37%; 蓝藻门在除了AQ4外的土样中普遍存在, 并且是AQ1的优势固氮菌群. 此外, 还发现了4个新的nifH基因, 分别是AQ1-12(KC412109)、 AQ4-3(KC412133)、 AQ4-4(KC412133)、 AQ4-5(KC412133), 其中后三者是土样AQ4的优势种群. 相似文献
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研究了久效磷( Monocrotophos,MCP)对秀丽隐杆线虫(Caenorhabditis elegans)胁迫相关基因mRNA表达的影响.结果表明久效磷暴露能够导致hsp-16.41、hsp-70、cyp-35a2、vit-2、vit-6和age-1基因表达量显著升高,cep-1、ape-1、sod-1、sod-3、ctl-2、ctl-3、gst-1、daf-12和daf-21基因表达量显著降低,对hsp-16.2、mtl-1和mtl-2基因表达量无显著影响.通过主成分分析发现,久效磷主要影响hsp-70、ctl-2、ctl-3、vit-2、cep-1和age-1基因表达,这些基因可作为久效磷毒性效应的敏感分子生物标志物. 相似文献
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Zosteramarina, a monocotyledonous angiosperm, is one of themost important seagrass species. To investigate the salt-tolerancemechanismand discover salt-tolerant genes in Z. marina, a cDNA library was constructed. Single-pass sequencing of the 5' ends of 4 081 clones yielded 4 002 high quality expressed sequence tags (ESTs), which were assembled into 241 contigs and 1 673 singletons, representing 1 914 unigenes. The average length of the ESTs was 582 bp, with sizes ranging from100-1 500 bp. Basic Local Alignment Search Tool (BLASTX) analysis revealed that 1 664 unigenes had significant homology to known genes in the National Center for Biotechnology Information (NCBI) non-redundant (nr) database (E-value≤5-10). Among them, the twomost abundant genes encoded metallothionein (157 ESTs) and chlorophyll a/b-binding protein (38 ESTs), accounting for 7.1% and 1.7% of the total ESTs, respectively. Using Kyoto Encyclopedia of Genes and Genomes (KEGG), 1 462 unigeneswere assigned to 1 161 pathways (E-value≤5-10). A total of 938 unigenes were assigned Gene Ontology (GO) terms based on the GO hierarchy analysis, and InterProScan searches recognized 1 003 InterPro families. Three genes formetallothionein in Z. marina that belonged to Class II was identified. Results of this study will improve understanding of the molecular mechanisms of saline tolerance in Z.marina. 相似文献
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《African Journal of Marine Science》2013,35(3):591-599
The genus Conus has over 500 species and is the most species-rich taxon of marine invertebrates. Based on mitochondrial DNA, this study focuses on the phylogenetics of Conus, particularly the pennaceus complex collected along the Mozambican coast. Phylogenetic trees based on both the 16S and the 12S ribosomal genes grossly revealed the same groupings: one that clustered the individuals from the north (Pemba, Nacala and Island of Mozambique), another that grouped the individuals from the central/southern coast (Pomene/Massinga and Inhambane) and a third that assembled the specimens from the central/southern islands (Bazaruto and Inhaca). The 16SrRNA-based trees further distinguished the northern group into a Nacala group and a Pemba+Island of Mozambique group. In all trees, C. p. bazarutensis and C. lohri collected on the central/southern islands grouped separately from the C. p. bazarutensis and C. lohri collected on the central/southern coast, suggesting a genetic similarity between these species. Likewise, C. praelatus revealed greater proximity to C. pennaceus from Pemba. Although different topologies were produced by each gene (low bootstrap support in some nodes), we support the hypothesis of a southward ancestral colonisation pattern, indicated by the 16SrRNA trees. The common ancestor would have shifted from planktonic to non-planktonic larval development and this weak vagility would have promoted the divergence between north and south specimens. Our results suggest that the separation of these groups might have been a relatively recent event, and part of the current morphological variability could be the outcome of phenotypic plasticity and/or ecological adaptation. 相似文献