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1.
建立黑斑原鮡(Glyptosternum maculatum)AFLP反应体系,对DNA提取、双酶切、连接、预扩增、选择性扩增和银染效果进行了探索。结果表明,40μLTaqI酶切体系中65℃先酶切2h,EcoRI酶切和连接同时进行,EcoRI酶和T4DNA连接酶的用量均以2U为宜,温度与时间分别为37℃和3h,连接产物稀释10倍作为预扩增的模板。利用这种优化的反应体系,5对选择性扩增引物组合表现出较高的稳定性、清晰度和多态性,它们分别是:E-AAC/T-AAT、E-AAC/T-AAG、E-AAG/T-ACA、E-AAG/T-AAT和E-AAT/T-AGA。5对引物组合对96个样本扩增,共得到332条清晰可辨、可重复的条带,平均每对引物组合扩增出66.4条带,其中51条带是多态的。将AFLP技术用于黑斑原鮡的研究,通过优化体系的建立,为其种质资源的研究奠定基础。  相似文献   

2.
以珠江长臀(Cranoglanis bouderius)为试验材料,建立了长臀扩增酶切片段长度多态性(AFLP)分析体系,对其分析过程中的DNA提取、双酶切、单酶切、连接、预扩增、选择性扩增、银染等步骤进行了检测,并对反应条件进行了优化。用PstI/TaqI、EcoRI/MseI等2组双酶切组合构建长臀的AFLP指纹图谱,条带清晰可辨,扩增信号强,得到清晰的长臀AFLP指纹图谱。  相似文献   

3.
大菱鲆AFLP分析体系的建立   总被引:5,自引:0,他引:5  
本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。  相似文献   

4.
程飞  叶卫  叶富良  刘波 《南方水产》2007,3(4):46-51
以珠江长臀(鮠)(Cranoglanis bouderius)为试验材料,建立了长臀(鮠)扩增酶切片段长度多态性(AFLP)分析体系,对其分析过程中的DNA提取、双酶切、单酶切、连接、预扩增、选择性扩增、银染等步骤进行了检测,并对反应条件进行了优化.用PstⅠ/TaqⅠ、EcoRⅠ/MseⅠ等2组双酶切组合构建长臀鲍的AFLP指纹图谱,条带清晰可辨,扩增信号强,得到清晰的长臀(鮠)AFLP指纹图谱.  相似文献   

5.
本研究构建了西伯利亚鲟(Acipenser baeri)扩增片段长度多态性(AFLP)分析体系,分析了DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤.用7对引物对24尾西伯利亚鲟进行了AFLP分析,每对引物扩增的位点数在24~39之间,共扩增出了234个位点,多态性位点为116个,多态性比例为...  相似文献   

6.
甘肃金鳟RAPD反应体系构建及优化   总被引:1,自引:0,他引:1  
以甘肃金鳟尾鳍为试验材料,提取基因组DNA。对影响甘肃金鳟RAPD扩增的重要参数进行了优化试验,包括模板DNA浓度、dNTP浓度、引物浓度、Taq酶、变性时间、退火温度等,建立了甘肃金鳟RAPD反应的最适体系。即在25μl反应体系中:模板DNA用量为4.0 ng/μl;Mg2 浓度为2.5mmol/L;dNTP浓度为0.5 mmol/L;引物浓度为0.5μmol/L;Taq酶用量为1 U;扩增程序:94℃预变性4 min,94℃变性45 s,36℃退火50 s,72℃延伸1 min,共35个循环,最后延伸10 min。  相似文献   

7.
本研究构建了西伯利亚鲟(Acipenser baeri)扩增片段长度多态性(AFLP)分析体系,分析了DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤。用7对引物对24尾西伯利亚鲟进行了AFLP分析,每对引物扩增的位点数在24~39之间,共扩增出了234个位点,多态性位点为116个,多态性比例为48.83%,平均Nei氏基因多样性指数H为0.4297,平均Shannon氏指数I为0.1233,个体间平均遗传距离为0.1872,表明西伯利亚鲟养殖群体的遗传多样性程度比较低,同时还构建了西伯利亚鲟的聚类图。  相似文献   

8.
以魁蚶为研究材料,建立了魁蚶的AFLP分子标记反应体系。对其分析过程中的酶切链接、预扩增、选择性扩增等试验条件进行了检测与筛选,得到了清晰的AFLP指纹图谱,为探讨魁蚶种群之间的遗传关系和遗传图谱的构建等方面提供了新的技术手段。  相似文献   

9.
以赣江野生青鱼(Mylopharyngodon piceus)基因组DNA为试验材料,研究Mg2+浓度、dNTP浓度、引物浓度和模板浓度4个参数对ISSR-PCR反应的影响,建立一套适合青鱼ISSR-PCR反应的最佳体系。结果表明,在25μL的反应体系中,Mg2+、dNTP、引物和模板4个参数的最适宜浓度分别为:3.0mmol/L、0.25mmol/L、0.4umol/L和30ng。PCR扩增程序为:94℃预变性5min,94℃30s,52℃45s,72℃2min,共38个循环,最后72℃继续延伸10min。  相似文献   

10.
用超分支滚环扩增法检出传染性脾肾坏死病毒。超分支滚环扩增法包括锁式探针的连接及超分支滚环扩增法扩增两部分,通过研究锁式探针的最佳连接体系和连接条件,并进行超分支滚环扩增法反应条件的优化,得出的本研究最适反应条件为:锁式探针在T4 DNA连接酶作用下37℃连接20 min,接下来的超分支滚环扩增法在Bst DNA聚合酶大片段作用下61℃反应40 min,即能够实现靶序列的有效检出。灵敏度试验表明,超分支滚环扩增法所能检测到的最低模板量接近1 copy。在4种病毒中进行的特异性试验表明,本方法能够保证对传染性脾肾坏死病的特异性检测。  相似文献   

11.
12.
中国介螅水母属二新种(丝螅水母目,介螅水母科)   总被引:1,自引:1,他引:0  
记述了台湾海峡及其邻近海区介螅水母科Hydractiniidae L.Agassiz,1862介螅水母属Hydractinia van Beneden,1841二新种,即螺旋介螅水母,新种Hydractinia spiralis sp. nov.和反曲介螅水母,新种Hydractinia recurvatus sp.nov.,兹将二新种主要鉴别特征描述于下:螺旋介螅水母,新种:伞无顶突;垂管长圆柱形,约为内伞腔深度2/3;口腕简单,末端具成束刺胞;生殖腺很大,呈椭圆形,几乎覆盖整个垂管间辐位,无水母芽;缘触手6条,4条主辐位,另2条仅在相对间辐位,基球延长锥状,同样大小,无眼点,触手长,近端2/3粗壮,远端1/3变细,具螺旋状刺胞。反曲介螅水母,新种:伞无顶突;垂管短小,呈方形;胃柄显著,约为垂管长度1/2;口有4个主辐位口唇,延长成向上反曲的短口腕,末端具成丛刺胞;4个卵圆形生殖腺,附在二个反曲口腕之间的垂管间辐位,无水母芽;缘触手16条,触手基球膨大,呈锥状,同样大小,整条触手具环状刺胞,末端略膨大,呈椭圆形。  相似文献   

13.
The aquaculture industry relies on federal investments to stimulate aquaculture research and development (R&D). We conducted this study to track federal spending for aquaculture using a large database of 2957 US federal research grants awarded from 1990 to 2015. During that period, 1.04 billion US dollars (USD) were spent on aquaculture research, with 919 million USD from federal agencies and 123 million USD in matching funds. By comparison, over the same time frame, the US Department of Agriculture's R&D spending was 41 billion USD. Federal grant funding for aquaculture had a 37‐fold return on investment since 2000. Microalgae, oysters, salmon and trout, catfish, and shrimp received the most grant support, and the most common topics/disciplines were aquaculture production sciences, aquatic animal health and disease, genetics and breeding, and nutrition. Our findings can be used to identify research gaps and inform federal policy, grant making, and budget allocation.  相似文献   

14.
In the present study, protective effects of dietary eucalyptol were investigated against copper toxicity in common carp (Cyprinus carpio). The fish were fed with diets supplemented with 0 (control), 0.5% and 1% eucalyptol for 14 days before exposure to 0.25 mg/L ambient copper for 7 days. Blood samples were taken from the fish before and 1, 3 and 7 days after copper challenge. Before copper challenge, eucalyptol significantly decrease serum aspartate aminotransferase (AST) activity, but increase blood WBC, RBC, hematocrit and haemoglobin, serum lysozyme, total immunoglobulin (Ig) compared with the control. Copper exposure led to stress, tissue damage and immunosuppression. After the copper challenge, significant elevation in serum cortisol, glucose, alanine aminotransferase (ALT) and AST, but decrease in blood RBC and WBC, and serum lysozyme, complement (ACH50) and total immunoglobulin (Ig) was observed. Eucalyptol administration significantly mitigated the increase in serum cortisol, glucose, ALT and AST, and decrease in WBC, ACH50, lysozyme and total Ig. Dietary 1% eucalyptol had significantly higher protective effects than 0.5% in the case of serum AST, lysozyme, total Ig and ACH50. In conclusion, eucalyptol has beneficial health effects on common carp and enables to mitigate copper‐induced stress, tissue damage, and immunosuppression. Eucalyptol at the dietary 1% level is recommended to protect common carp against adverse effects of copper toxicity.  相似文献   

15.
采用L25(56)正交试验法测定了不同pH(6、7、8、9、10)、温度(10、15、20、25、30℃)和盐度(15、20、25、30、35)对体质量为(71.2±8.5)g的单环刺螠肠蛋白酶、淀粉酶、纤维素酶和脂肪酶活力的影响,以及不同盐度对单环刺螠血液溶菌酶活力的影响。试验结果表明,单环刺螠肠消化酶活力的最适环境条件分别为,蛋白酶:pH 8、温度30℃、盐度35;淀粉酶:pH 9、温度25℃、盐度25;纤维素酶:pH6、温度30℃、盐度35;脂肪酶:pH 6、温度25℃、盐度30。盐度15和20两组单环刺螠血液溶菌酶活力均先降后升再降,而盐度30和35两组则先升后降。至处理第4d,各试验组溶菌酶活力依次为:盐度3025352015,盐度15和20两组溶菌酶活力显著低于盐度较高试验组(P0.05)。盐度25~35、pH 6~9为单环刺螠适宜的环境条件,高温(25~30℃)下其消化酶活力较高,而低盐度(15~20)下其消化酶活力和免疫能力明显降低。  相似文献   

16.
The northeast region of India, comprised of the states of Arunachal Pradesh, Assam, Manipur, Meghalaya, Mizoram, Nagaland, Sikkim, and Tripura, is blessed with rich biodiversity and fisheries resources. With more than 90% of population being fish eaters, there is heavy demand for fish but a wide gap exists between supply and demand. The region produces over 0.214 million tons of fish annually, with almost 50% coming from aquaculture. Aquaculture development in the region is taking place at a rapid rate. However, efforts are necessary to increase the present level of production through both horizontal and vertical expansion. The region has rivers, coldwater streams, floodplain wet lands, reservoirs, lakes, ponds, paddy fields, and mini-barrages to support large-scale aquaculture activities, which can not only produce fish to meet regional requirements, but also export the surplus. This paper describes and discusses the current status of aquaculture, production level, constraints, and future directions towards achieving “fish for all” in the region.  相似文献   

17.
运用线粒体D-loop区与COI基因片段序列比较分析了养殖与野生银鲳群体的遗传多样性。研究结果显示,线粒体D-loop区片段中,A、T、C与G4种核苷酸的平均含量分别为40.00%、30.55%、16.75%和12.70%,A+T的含量为70.55%,明显高于G+C的含量。COI基因片段中,A、T、C和G的平均含量分别为25.85%、33.90%、21.30%和18.85%,A+T的含量(59.75%)同样高于G+C的含量。基于D-loop序列分析所得出的两群体总的变异位点、单倍型数、单倍型多样性、核苷酸多样性及平均核苷酸差异数分别为19、15、0.895、0.007和2.505。基于COI基因所得出的两群体总的变异位点、单倍型数、单倍型多样性、核苷酸多样性及平均核苷酸差异数分别为33、17、0.713、0.004和2.239。基于线粒体D-loop区与COI基因片段序列的研究结果均显示,养殖银鲳群体的遗传多样性低于野生群体的遗传多样性。养殖群体基于线粒体D-loop区与COI基因片段分析得出的单倍型多样性分别为0.562与0.571,野生群体基于线粒体D-loop区与COI基因片段分析得出的单倍型多样性分别...  相似文献   

18.
The objective of the present study is to preliminarily clarify the mechanism of carbohydrates metabolism in cobia (Rachycentron canadum) (85 ± 3 g) receiving injection of glucose solution. We examined plasma glucose (GLU), total protein (TP), triglyceride (TG), cholesterol (CHOL), insulin, liver glycogen and muscle glycogen, activities of hepatic hexokinase (HK), phosphofructokinase (PFK) and pyruvate kinase (PK), as well as relative expressions of glucose transporter 1 (GLUT1), GLUT2, GLUT3, GLUT4, GLUT5 and GLUT9 in hemocyte, liver and muscle of R. canadum when fish were injected with 200 μl of glucose solution (255 mg/ml) after 0, 1, 2, 4, 8, 12, 24 and 48 hr. Fish received injection of 0.68% saline served as control. Results indicated that the plasma GLU, TG and CHOL increased and reached peak at 1, 8 and 48 hr postinjection (hpi) respectively. The hepatic glycogen increased from 1 hpi, and reached peak at 8 hpi, plasma insulin increased at 1 hpi, and reached peak at 2 hpi, and activity of hepatic PK peaked at 8 hpi. Furthermore, the relative expressions of GLUT1, GLUT2, GLUT3, GLUT4 and GLUT5 in hemocytes reached peak at 1,4, 8, 4 and 8 hpi, respectively, relative expressions of GLUT2, GLUT3, GLUT5 and GLUT9 in liver reached peak at 24, 24, 12 and 24 hpi, respectively, and relative expressions of GLUT1 and GLUT3 in muscle were significantly higher at 2 and 2–4 hpi, respectively compared with those in controls. In conclusion, low ability of utilizing glucose in R. canadum may be attributed to insufficient insulin secretion, low activities of key glycolytic enzymes (HK, PFK and PK) regulated by glucose injection and slow increase of GLUTs.  相似文献   

19.
Data sets with increased spatial and temporal resolution can help researchers and resource managers quantify representative distributional patterns of mobile sportfish. In this research, first, we illustrate patterns of sportfish distribution using individual (percent of population, residence time, number of movements) and combined distributional metrics. Second, we apply these metrics to one highly mobile fish species, the blue catfish (Ictalurus furcatus), across a range of spatial (whole reservoir, region, site) and temporal (year, month, diel period) scales. Specifically, we tracked 123 acoustically tagged blue catfish with a 20‐receiver array in Milford Reservoir, KS, USA. When we integrated metrics, four site‐specific distributional patterns emerged: (a) a large, active multi‐site fish aggregation, (b) localised site fidelity, (c) transitional sites and (d) rarely used locations. These patterns would not have been detected using a single metric as each measurement revealed a different piece of the distribution story. For example, if we had only quantified percent of population, we could identify fish location, but not whether individual fish spent time at a location or were just passing through. Our examination of multiple scales also provided a novel context for interpreting site‐specific patterns. As an illustration of this insight, using conventional approaches, we would have observed heterogeneity, but we would not have detected fish aggregations, in which individual fish either remained or repeatedly returned to a site. In summary, our results show the advantage of setting the entire ecosystem as the study boundary to integrate multiple responses using a spatially and temporally extensive data set.  相似文献   

20.
The tropical gar (Atractosteus tropicus) is an economically and socially important freshwater fish species from southeastern Mexico, with high aquaculture potential. This study determined the expression of acetyl‐CoA carboxylase (ACC1), fatty acid synthase (FAS), and carnitine palmitoyl transferase 1 (CPT1C) in several tissues of adult specimens, as well as in early ontogeny stages of tropical gar. Partial sequences of all genes were obtained for brain, liver, muscle, mesenteric adipose, kidney, testis, gill, and heart in males. Specific qPCR primers were used to compare the expression between tissues and also during larviculture, starting with embryos (0 days after hatching, [DAH]), and at 5, 10, 15, 20, 25, and 30 DAH. Mesenteric adipose tissue, liver, and muscle showed maximum FAS, ACC1, and CPT1C expression. Additionally, the highest expression of lipogenic genes was detected in embryos (0 DAH), decreasing between 5 and 10 DAH, then increasing at 15 DAH, and ultimately decreasing again from 20 DAH onwards. Based on these results, it can be concluded that genes encoding ACC1, FAS, and CPT1C are expressed differentially across tissues of tropical gar adults, and are regulated during larval development.  相似文献   

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