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中国鲎(Tachypleus tridentatus)基因工程抗菌肽的制备及其抗菌活性 总被引：3，自引：0，他引：3
从中国鲎血细胞RT-PCR扩增抗菌肽tachyplesins基因,经改造构建原核和真核表达载体pET-KRN、pPIC9-KRY,并分别在大肠杆菌DE3和毕赤酵母GSll5中诱导表达.用Tris-Tricine SDS-PAGE检测并进一步电洗脱纯化表达产物,经体外生物活性测定表明,表达产物KRN对大肠杆菌、金黄色葡萄球菌、枯草杆菌、伤寒沙门菌的抑菌环为25、22、25、16mm;MIC值为3.57、7.14、3.57、14.28mg/L.表达产物KRY对大肠杆菌、金黄色葡萄球菌、枯草杆菌、伤寒沙门菌和李氏杆菌的抑菌环为26、22、25、18、16mm;MIC值为1.57、3.14、1.57、6.28、6.28mg/L,对水霉的抑菌效果尤其显著,抑菌环达35mm. 相似文献
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采用正交设计法,从dNTPs浓度、引物浓度、Mg2＋浓度、Taq DNA聚合酶用量4个因素3个水平出发,优化设计圆尾鲎DNA的PCR反应体系（引物为中国鲎微卫星引物）.并采用直观分析方法分析正交试验结果,最终建立了圆尾鲎SSR-PCR最佳反应体系：总体积20μL,Taq DNA聚合酶1.5 U、dNTPs 0.16 mmol/L、引物0.2μmol/L、Mg2＋2.0 mmol/L;并通过PCR梯度实验进一步优化模板DNA质量浓度、退火温度及退火时间,获得最佳反应条件：模板DNA质量浓度为30 ng/μL,退火温度为48℃,退火时间为20～25 s.对最佳反应体系和反应条件进行了检验,结果显示该反应体系稳定性高、重复性好. 相似文献
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Kit Yue KWAN Justin BOPP Shuyan HUANG Qiao CHEN Chun-Chieh WANG Xueping WANG Wenquan ZHEN Junhua ZHU Xing HUANG 《Integrative zoology》2021,16(6):908-928
Horseshoe crabs, the most well-known example of “living fossils”, are iconic and ecologically important macroinvertebrates in coastal and estuarine ecosystems. Their blood is a crucial resource for manufacturing Limulus or Tachypleus amebocyte lysate to detect bacterial endotoxins or fungal contamination in drug and medical devices. An enhanced understanding of their ecological roles and trophic interactions in the food webs is fundamental to facilitate resource management for the declining populations in Asia. Foraging information of the Asian species, however, is mainly derived from preliminary, scattered reports from a limited number of study locations. In this study, resource utilization, trophic niche dynamics, and trophic interaction of the juvenile tri-spine horseshoe crab, Tachypleus tridentatus (instars 1–12, approximately 0.5–8 years old) across ontogeny was assessed in diversified nursery habitats along the northern Beibu Gulf, China, using carbon and nitrogen stable isotopes. Our results suggest that: (i) T. tridentatus are ecological generalists given the vast range of carbon isotopic values and trophic niche width estimates exhibited between multiple instar groups; (ii) juvenile T. tridentatus across most habitat types predominantly assimilated energy from a variety of basal production sources in the food web, but primarily depended on sedimentary organic matter and seagrass resource pools; (iii) ontogenetic shifts in juvenile dietary proportions were evident, with decreased reliance on sedimentary organic matter, coupled with increased reliance on benthic macroinvertebrate grazers, detritivores, and omnivores with age; and (iv) nearly all juvenile instars occupied similar trophic positions in the food web with slight shifts in trophic position present with increasing size. Our findings indicate that resource availability and ontogenetic diet shifts strongly influence horseshoe crab trophic dynamics, and age should be accounted when formulating habitat conservation measures based on resource use for Asian horseshoe crabs. 相似文献
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应用等位酶技术对漳浦、连江和温州3个地理群体的中国鲎遗传多态性进行分析.实验结果发现：6个酶系统的10个位点中有6个为多态性位点,共获得21个等位基因;大部分位点在3个群体中偏离Hardy-Weinberg平衡;在物种水平上,有效等位基因数为1.635,观察杂合度为0.456,期望杂合度为0.306,香侬指数为0.486,表明中国鲎的遗传多样性较高;F-统计量（FST）平均为0.0475,表明中国鲎遗传差异主要存在于群体内;遗传一致度平均为0.969,遗传距离平均为0.0315,表明中国鲎群体间遗传分化较低;基因流平均为5.0120,显示群体间存在较大的基因交流.由此认为3个群体属于同一个繁殖群体.在进行鲎物种保护的同时,要加强对其遗传多样性的保护. 相似文献
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鲎C因子是鲎血细胞中一种对内毒素具有高亲和力的丝氨酸蛋白酶原,可替代鲎试剂用于内毒素检测。鲎C因子N端Cys—EGF．sushil—sushi2（CES）片段中的sushil区域是与内毒素结合的关键部位。本研究在sushil结构域中的特定位点断开CES与内毒素结合的功能片段,然后分别和蛋白质内含肽SspDnaX的N端片段（IN）和C端片段0C）连接,并在大肠杆菌中表达,表达产物经亲和层析纯化、复性以及内毒素去除后,利用蛋白质反式剪接系统,在体外诱导该蛋白质内含肽发生剪接,重新得到C因子的CES蛋白,并检测其活性。实验结果表明此重组CES蛋白具有结合内毒素活性,提示在大肠杆菌系统中开发低成本内毒素检测试剂的可行性。 相似文献
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为了探究不同龄期中国鲎幼鲎生长特性,实验测定了2～4龄幼鲎头胸部长(X1)、头胸部宽(X2)、腹部长(X3)、腹部宽(X4)、剑尾长(X5)、全长(X6)和体重(Y)等参数,运用相关分析和通径分析方法,探讨形态性状对体重的作用效果。结果显示,2～4龄幼鲎体重相关系数最高的形态性状均为头胸部宽(X2);通径分析发现,头胸部宽(X2)对2龄和3龄幼鲎体重直接影响最大,腹部宽(X4)对4龄幼鲎体重直接影响最大;各形态性状对体重决定程度分析与通径分析变化趋势一致。对2～4龄幼鲎头胸部宽和体重进行对数处理,获得异速生长方程回归系数(b)为2.746,表明此阶段幼鲎体重增长率大于头胸部宽增长率,处于正生长期。实验表明,不同龄期阶段中国鲎苗种筛选所依据的形态性状有所差异,中国鲎幼鲎阶段种质评价应考虑其异速生长规律,同时也为探究鲎的生长与性状、结构与生理之间的关系奠定基础。 相似文献
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Marine microalgae as dietary supplements in the culture of juvenile Chinese horseshoe crabs,Tachypleus tridentatus (Xiphosura) 
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下载免费PDF全文 Billy K Y Kwan Alice K Y Chan Siu Gin Cheung Paul K S Shin 《Aquaculture Research》2017,48(7):3910-3924
Five treatments (short‐neck clam Ruditapes philippinarum and four formulated diets with the green microalgae Dunaliella tertiolecta or the golden‐brown microalgae Isochrysis galbana in the ratio (dw/dw) of 5% or 10% to that of clam meat) were used to investigate the growth performance and haemolymph quality of juvenile Chinese horseshoe crabs Tachypleus tridentatus under laboratory culture. At the end of the 12‐week study, various growth and moulting indicators as well as haemolymph biochemical parameters did not show any significant differences among the treatments. Significantly higher percentages of haemolymph amoebocyte viability and granular‐spherical amoebocyte state were noted in both 5% and 10% I. galbana‐supplemented diets compared to that of clam meat. However, the percentages of these two haemolymph parameters in juveniles fed with D. tertiolecta decreased significantly at the end of the experiment. Such differences were attributed to the presence of polyunsaturated fatty acid (PUFA), notably eicosapentaenoic acid (EPA), in I. galbana that was absent in D. tertiolecta and clam meat. Like many marine invertebrates, juvenile horseshoe crabs do not naturally obtain sufficient PUFAs from their diets to meet developmental and physiological needs; hence, supplementing microalgae with high levels of EPA in a protein‐rich diet boosts immune competence and the health status of juvenile horseshoe crabs while under culture. 相似文献
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检测分析成年雌性中国鲎不同组织的营养成分,探索其繁殖期的营养需求,寻找促进中国鲎性腺发育和卵细胞成熟的方法。采用国标生化分析方法对成年雌性中国鲎肌肉、黄色结缔组织和卵的常规营养、氨基酸和脂肪酸进行测定。结果表明,3种组织在常规营养成分、氨基酸和脂肪酸组成上较为一致,但在含量上存在着明显差异,肌肉水分含量最高(77.5%),卵粗蛋白含量最高(27.9%),黄色结缔组织粗脂肪(16.5%)、粗灰分(1.6%)、钙(1220 mg/kg)、磷(107 mg/100 g)含量均最高。3种组织中检测出17种氨基酸、13种脂肪酸;氨基酸组成中以谷氨酸(Glu)含量最高,脂肪酸组成中以棕榈酸(C16:0)和油酸(C18:1)含量为最高。DHA和EPA含量均为卵>黄色结缔组织>肌肉。以上结果说明,成年雌性中国鲎肌肉具有高蛋白质、低脂肪的特点,卵中含有丰富的氨基酸和不饱和脂肪酸。研究结果对于其种质资源保护、合理开发利用和营养价值评价均具有一定参考意义。 相似文献
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应用光镜及电镜技术研究中国鲎（Tachypleus tridentatus（Leach））复眼的形态和超微结构。结果表明，中国鲎复眼表面覆盖角膜层，外形呈半球形，由800-1200个小眼组成，组成复眼的小眼面均为椭圆形。每个小眼包括透光和感光两部分。透光部分由透光锥体和透光孔径组成；感光部分包括小网膜细胞及色素细胞。小网膜细胞分R-区和A-区，R-区是有感杆束分布的区域，A-区是没有感杆束分布的区域，但A区的表面覆盖着色素细胞，色素颗粒主要分布于A-区和R-区的交界处。8个小网膜细胞排列成的桔瓣状结构组成1个小眼的感光部分。[中国水产科学，2006，13（4）：517—524] 相似文献