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1.
基于cox1片段序列的DNA条形码为浮游动物种类鉴定提供了新的可靠手段,构建了胶州湾网采浮游动物DNA条形码文库,在国内首次使用宏遗传组学方法研究夏季胶州湾海洋浮游动物的群落组成。共获得环境样品DNA条形码149条;以6%为阈值共检出37个OUT,其中19个与DNA条形码数据库中序列程度非常高(〉97%),可以鉴定到种...  相似文献   

2.
A cancer-resistant fish, the mummichog Fundulus heteroclitus, was assayed for its ability to repair damage to its genome caused by an alkylating agent. Although DNA may be alkylated at several sites, the persistence of O6-alkylguanine has been correlated with carcinogenesis. O6-alkylguanine, if not repaired before the next round of replication, can lead to base transition mutations. The DNA repair protein, O6-alkylguanine transferase (O6-AGT), removes alkyl groups from the O6 position of guanine.Populations from polluted and relatively pristine estuaries were compared. O6-AGT activity was assayed by incubating extracts of liver from feral fish with previously methylated calf thymus DNA. After incubation, the DNA was depurinated by acid hydrolysis and the O6-methylguanine (O6-MeG) was separated and quantified by HPLC. Activity was measured by the reduction of O6-MeG in the incubated sample as compared to controls.Both populations demonstrated O6-AGT activity in their livers. Both had activity levels which were not as high as those seen in mammals. The reference population had O6-AGT levels inversely correlated with size, which is expected since younger fish have more DNA synthesis in their livers. The polluted population did not have a significant correlation with size, and generally exhibited less activity than the reference population. The lower O6-AGT activity in the polluted population suggests an environmental impact, and does not answer the question as to why this species is cancer resistant.  相似文献   

3.
Reproduction by sexual or asexual viviparity is a common phenomenon in some anemone species. In this short communication, the origin of the brooded young of Actinia equina and A. schmidti from the Portuguese shore was investigated. DNA was extracted from 56 brooding adult Actinia sp. and the nuclear gene that codes for the 28S ribosomal subunit was sequenced. Species identity was then assessed using GenBank. In total, 50 individuals were A. schmidti, five were A. equina and one had a hybrid origin. Three adult anemones (the hybrid, one A. equina and one A. schmidti) possessed two different 28S sequences and so their offspring was selected for further analysis using the same molecular procedure. Each brooded polyp was found to possess the exact same sequence as its parent, strongly suggesting the asexual origin of broods in A. equina and A. schmidti.  相似文献   

4.
A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid detection of the toxic microalgae Alexandrium catenella and A.minutum,which can produce paralytic shellfish poisoning (PSP).Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA.The method worked well in less than an hour under isothermal conditions of 65 C.LAMP specificity was validated in closely related algae as a comparison,suggesting the strict specificity of the LAMP primers.Two visual inspection approaches were feasible to interpret the positive or negative results.The detection limits of A.catenella and A.minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA,respectively.The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae.These characteristics of species specificity,sensitivity,and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A.catenella and A.minutum.  相似文献   

5.
为检测中国近海海洋环境中菊酯类农药等污染物对日本囊对虾(Marsupenaeus japonicas)的基因毒性及其危害程度,作者首次使用醚菊酯对日本囊对虾进行毒理实验,将日本囊对虾分为6组进行不同剂量的处理,采用彗星实验分析技术对处理培育25 d的日本囊对虾DNA损伤情况进行检测,并通过Comet Score 1.5分析软件对实验图谱进行拖尾率、尾长、彗尾DNA相对含量、尾距、Olive距等DNA损伤指标的统计分析,在对各指标分别建立一元回归方程的基础上,又通过SPSS软件进行判定分析,筛选影响作用更大的检测指标,建立了多参数的多元回归方程。实验结果发现,与空白对照组相比,日本囊对虾各实验处理组肝胰腺细胞DNA均有不同程度的损伤且各检测指标均有显著性差异(P0.01),各检测指标与处理浓度表现出良好的剂量效应关系,随着处理浓度递增,各检测指标都呈规律性的增长趋势。回归分析和相关系数表明:各检测指标与处理浓度具有高度的相关性,多项式回归方程(P0.05)和多元回归方程(P0.005)均有显著性的统计学意义,通过多元回归方程可以有效地推断醚菊酯处理浓度和处理时间。  相似文献   

6.
Laminaria (Saccharina) is one of the economic important seaweeds.Through sequence analysis of 4 099 ESTs from Laminaria (971 were generated from our own cDNA laboratories and 3 128 were downloaded from updated EST databases) with SSRIT software,two hundred and fifty-four SSRs in 206 ESTs were found.From the 254 SSRs,sixty-three SSR primer-pairs were designed. In order to test their practicability, the 63 primer-pairs were tested in commonly used SSR reaction conditions using 12 Laminara DNA samples as templates. The results show that 23 SSR primer-pairs gave good amplification patterns on most (more than 80%) of the 12 Laminara DNA templates. Genetic diversity study of 12 Laminaria lines,which were widely used in breeding and economic cultivation in China,was performed based on the obtained SSR data.  相似文献   

7.
为了阐明南中国海裸胸鳝属(Gymnothorax)鱼类系统进化情况,利用聚合酶链式反应扩增目的产物,将其连接到T载体后,并对其序列进行测定,共得到9种裸胸鳝属鱼类线粒体16S ribosomalDNA(16S rDNA)基因的部分序列,采用多个生物软件对序列变异和碱基组成进行分析,计算了Kimura2-parameter遗传距离,转换/颠换比等遗传信息指数,下载基因库中16SrDNA基因的同源序列,以鳗鲡属(Anguilla)的花鳗鲡(Anguilla marmorata)为外群构建NJ(Neighbore-Joining)、MP(MaximumParsimony)和ME(Minimum Evolution)系统进化树。根据所得分子数据并结合形态学特征推论如下:(1)在所研究的10种裸胸鳝鱼中共有516个位点、其中143个核苷酸位点存在变异(27.7%);(2)序列变异的转换/颠换比值的平均值为3.441;相对遗传距离数据表明斑颈裸胸鳝(G. margaritophorus)和网纹裸胸鳝(G. reticularis)差异最大(0.177),褐祼胸鳝(G. hepaticus)与布雷顿氏裸胸鳝(G. breedeni)差异最小(0.022);(3)NJ树、ME树表明裸胸鳝属内部存在3个平行进化的姐妹分支,分支内部的种类组成与地理分布无关。  相似文献   

8.
Pfiesteria shumwayae Steidinger et Burkholder is now known to be present in New Zealand and occurs in estuaries around the country. The presence of Pfiesteria was initially determined by a polymerase chain reaction (PCR)‐based detection assay, using oligonucleotide primers targeted at ribosomal DNA extracted from estuarine water and sediments. Presence was confirmed by isolation from fresh sediments in the presence offish (Oreochromis mossambicus), followed by identification by scanning electron microscopy. The New Zealand isolates of P. shumwayae were ichthyotoxic in bioassays, but there is no historic evidence offish kills in New Zealand associated with the dinoflagellate.  相似文献   

9.
The in vivo response of freshwater fish exposed to pollutants was assessed using two biomarkers, 7-ethoxyresorufin-O-deethylase (EROD) induction and DNA single strand breaks. Chub (Leuciscus cephalus) were caught in spring and in fall at various locations in the river Rhône watershed. EROD activity was measured in the liver while DNA damage was evaluated in chub erythrocytes using the recently developed Comet assay. Chemical contamination was evaluated both in fish muscle (PCBs) and in sediment (PCBs, PAHs, heavy metals) collected at each sampling station. Sex of individuals was shown to influence the level of EROD activity but not the level of DNA damage. The EROD activity as well as the DNA damage were found to be higher in the mostly contaminated stations compared to the reference one. This study shows that multibiomarker-based approach provides complementary informations about early effects in feral fish exposed to complex chemical pollution and highlights the interest of the Comet assay in genotoxicity assessment.  相似文献   

10.
Multiple paternity was assessed in Octopus minor using microsatellite DNA markers. Octopus minor adults were captured in traps and kept in indoor cement ponds. The females spawned after several days. Ten broods (B1–B10), each containing 15 embryos and the maternal octopus, were maintained until the embryos reached the stage at which DNA could be extracted and used for genotyping. Multiple paternity was tested using five microsatellite DNA markers and the results proved the hypothesis that multiple paternity occurs in O. minor (observed in six out of the 10 broods). Analysis with GERUD software gave the minimum number of fathers and showed that four broods were sired by a single male, three were sired by two different males and three were sired by three different males. In brood B10, the number of fathers identified by the single‐locus method was one fewer than that by the GERUD method. Of the six multiple paternity broods, four (66.7%) showed significant skew from equal paternal contributions, which indicated that sperm competition and/or cryptic female choice may be important for post‐copulatory paternity bias in this species.  相似文献   

11.
We applied DNA‐based faecal analysis to determine the diet of female Australian sea lions (n = 12) from two breeding colonies in South Australia. DNA dietary components of fish and cephalopods were amplified using the polymerase chain reaction and mitochondrial DNA primers targeting the short (~100 base pair) section of the 16S gene region. Prey diversity was determined by sequencing ~50 amplicons generated from clone libraries developed for each individual. Faecal DNA was also combined and cloned from multiple individuals at each colony and fish diversity determined. Diets varied between individuals and sites. Overall, DNA analysis identified a broad diversity of prey comprising 23 fish and five cephalopod taxa, including many species not previously described as prey of the Australian sea lion. Labridae (wrasse), Monacanthidae (leatherjackets) and Mullidae (goat fish) were important fish prey taxa. Commonly identified cephalopods were Octopodidae (octopus), Loliginidae (calamary squid) and Sepiidae (cuttlefish). Comparisons of fish prey diversity determined by pooling faecal DNA from several samples provided a reasonable but incomplete resemblance (55–71%) to the total fish diversity identified across individual diets at each site. Interpretation of diet based on the recovery of prey hard‐parts identified one cephalopod beak (Octopus sp.) and one fish otolith (Parapriacanthus elongatus). The present study highlights the value of DNA‐based analyses and their capabilities to enhance information of trophic interactions.  相似文献   

12.
13.
The nitrogenase activities of cyanobacteria of the genus Trichodesmium are strictly light-dependent, although they do not develop heterocysts. Recently, the development of heterocyst-equivalent cells (diazocytes) was suggested in Trichodesmium spp. However, no cells with a similar appearance to diazocytes could be found in Trichodesmium sp. NIBB1067. An immunocytochemical analysis of nitrogenase in individual cells was performed using polyclonal antibodies generated from DNA fragments of genes encoding the Fe-protein and the α-and the β-subunit of the MoFe-protein of nitrogenase of Trichodesmium sp. NIBB1067. Visualization of the antibody binding was carried out using a horseradish peroxidase-conjugated secondary antibody and a chromogenic substrate, 3,3′-diaminobenzidine, to avoid the masking effects caused by the bright autofluorescence emitted by Trichodesmium cells. Nitrogenase proteins were detected in almost all the cells (always higher than 95% of total cells) grown under nitrogen-limited conditions. These results indicate that Trichodesmium sp. NIBB1067 does not differentiate heterocyst-equivalent cells.  相似文献   

14.
6种舌鳎亚科鱼类ITS1序列长度多态性及系统分析   总被引:4,自引:0,他引:4       下载免费PDF全文
利用核糖体第一内转录间隔区(ITS1)对舌鳎亚科(Cynoglossinae)6种鱼类进行系统分析,发现舌鳎亚科ITS1区具有明显的序列长度多态性(404—744bp),序列长度分为三种类型,分别为404—405bp、462—463bp以及741—744bp,同一长度类型的不同种类间序列高度相似,平均遗传距离分别为0.00248、0.00217和0.00169,而不同类型间序列差异显著,平均遗传距离最小为0.38453。分析表明,序列长度多态性可能与物种分化时间有关。采用NJ(neighbour-joining)法及MP(maximum parsimony)法构建分子系统树,结合形态学特征及GenBank中的线粒体DNA序列进行分析,表明紫斑舌鳎(Cynoglossus purpureomaculatus)与短吻三线舌鳎(C.abbreviatus)可能为同物异名。另外,舌鳎属(Cynoglossus)的中华舌鳎(C.sinicus)与须鳎属(Paraplagusia)的日本须鳎(Paraplagusia japonica)聚为一支,舌鳎属中三线舌鳎亚属(Areliscus)的长吻红舌鳎(C.lighti)与拟舌鳎亚属(Cynoglossoides)的少鳞舌鳎(C.oligolepis)聚为一支,与形态分类学的观点不一致,值得进一步研究。  相似文献   

15.
用基因枪将GUS基因导入褐藻细胞中表达   总被引:18,自引:2,他引:18  
于1993年11月-1994年2月,用高压氦气式基因枪,将PH1221质粒装有GaMV 35S启动子,GUS基因以及nos的3'调控区导入海带和裙带菜组织切块中。48h后,在海带假根细胞和裙带菜中肋部叶片细胞中检测到GUS基因的表达。实验表明,微粒子轰击法是外源基因导入大型褐藻的一个有效途径。GaMV 35S启动子能够驱动外源基因在海洋藻类中的表达,可作为藻类基因工程的启动元件。  相似文献   

16.
基于线粒体cox1片段序列的胶州湾浮游动物DNA条形码分析   总被引:3,自引:0,他引:3  
采用cox1基因特异扩增测序的方法,分析了胶州湾45种常见海洋动物的DNA条形码序列82条,联合GenBank中28条cox1序列的分析结果表明:种内个体间遗传差异均值为0.013(0—0.11);属内不同种间遗传差异均值为0.265(0.137—0.369),是种内遗传差异的20多倍,条形码间隙明显。在分子系统树中,...  相似文献   

17.
为探讨虾夷扇贝(Patinopecten yessoensis)性别决定和雌雄同体形成的分子机制,利用兼并引物,以闭壳肌基因组DNA为实验材料,扩增和克隆了虾夷扇贝Dmrt基因的DM结构域,雌性、雄性和雌雄同体3种性别共获得2个具有不同DM序列的克隆,序列长度均为141bp,分别命名为Py Dmrt3和Py Dmrt4。Py Dmrt3在3种性别类型中均有克隆,而雄性中还克隆出Py Dmrt4。结果表明,在虾夷扇贝不同性别中,Dmrt基因家族的成员可能会有不同;雄性py Dmrt3的DM结构域核苷酸和氨基酸变异频率高,其次是雌雄同体,雌性的较保守,与其他软体动物DM结构域比对,该基因家族在进化上仍然高度保守,由此推测,部分雄性可能是发生了性逆转的雌雄同体,这种较高的变异性可能也是雌雄同体的一个特点,Py Dmrt4可能参与调控雄性性别的形成。  相似文献   

18.
微绿球藻(Nannochloropsis oculata)DNA被提取纯化并经超声波处理后,将所得大小在1.6~3 kb之间的DNA片段用T4 DNA聚合酶补平,再与SmaI酶切消化并经去磷酸化处理的质粒载体pUC18连接,转化至DH10B大肠杆菌(Escherichia coli)的感受态细胞中。建立的DNA质粒文库容量含2×104个克隆,其中重组子占90%。随机挑选白色菌落并培养,抽提的质粒经XbaI和SacI双酶切鉴定,显示重组的质粒中均含有大小不等的DNA插入片段。  相似文献   

19.
Turbot (Scophthalmus maximus) and mussel (Mytilus edulis) microsomes were incubated with DNA to examine if microsomal in vitro metabolism of BaP could result in DNA adducts detected by 32P-postlabelling. Turbot DNA was incubated with benzo[a]pyrene (BaP), NADPH and microsomal activating systems prepared from either livers of unexposed turbot, turbot exposed to BaP or β-naphthoflavone (ß-NF) or digestive glands from mussels. The β-NF activating system generated the highest levels of DNA adducts detected in this study (451.7 adducts per 108 nucleotides) and were distributed in three discrete adduct TLC spots, one of which (97% of the total adducts) co-migrated with the 32P-postlabelled BaP 7,8-diol, 9,10-epoxide-N2-guanine adduct. Fewer adducts (P <0.05) were generated by BaP-induced microsomes (9.4–30.6 adducts per 108 nucleotides) but levels were higher (P <0.05) than those generated from untreated fish (3.5 adducts per 108 nucleotides). Co-incubation with 500 μM α-naphthoflavone (α-NF) resulted in 97–99% inhibition in adduct formation implicating cytochrome P450-dependent (CYP) bioactivation however there was some evidence for carry over of BaP in the liver microsomal preparations from BaP injected fish. In contrast to the fish activating systems, no DNA adducts were observed when mussel microsomes were incubated with BaP, DNA and NADPH.  相似文献   

20.
选取12个微卫星标记对洞庭青鲫(Carassius auratus var.Dongtingking)、野生二倍体和三倍体鲫(C.auratus)、彭泽鲫(C.auratus var.pengzesis)4个鲫品系群体进行遗传多样性检测。在12个基因座位中,共检测出78个等位基因,其中14个为共有等位基因;每个座位检测到等位基因4—12个,平均等位基因数6.58个;4个鲫品系群体的平均观测杂合度(Ho)分别为0.633、0.750、0.800、0.717;平均期望杂合度(He)分别为0.502、0.713、0.757、0.602;平均多态信息含量(PIC)分别为0.364、0.599、0.637、0.470。上述结果表明,野生二倍体和三倍体鲫的遗传多样性较为丰富,以三倍体鲫的遗传多样性为最高;而洞庭青鲫和彭泽鲫养殖群体存在杂合度降低,遗传多样性下降的现象,以洞庭青鲫的遗传多样性为最低。基于遗传距离构建的UPGMA聚类树表明,洞庭青鲫与彭泽鲫两个养殖群体聚为一支,而二倍体与三倍体野鲫群体聚为另一支,说明洞庭青鲫与彭泽鲫之间亲缘关系较近,野生二倍体与三倍体鲫之间的亲缘关系较近。研究不同倍性鲫品系的遗传多样性,对于探讨鲫的多倍体起源演化以及鲫品系的种质资源保护和选育种实践具有重要意义。  相似文献   

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