共查询到15条相似文献，搜索用时 406 毫秒
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丝毛乌骨鸡胸肌和肝脏cDNA文库的构建与鉴定 总被引：1，自引：0，他引：1
了解肌肉组织和肝脏组织的基因类型,构建鸡肌肉组织和肝脏组织的功能基因表达谱在动物遗传育种中具有重要的意义.为比较基因组学的研究、功能基因组学的研究、QTLs定位等的动物分子育种做前期基础科研工作,研究以丝毛乌骨鸡(Gallus gallus)的胸肌和肝脏为实验材料,以Lambda ZAPⅡ为载体,构建了胸肌和肝脏的cDNA文库.同时,对这两个cDNA文库进行了噬菌体PCR鉴定和所提质粒的EcoRⅠ和XhoⅠ的双酶切鉴定.结果表明,胸肌和肝脏mRNA的OD260/OD280分别为1.92和1.90.胸肌cDNA文库的滴度在3.6× 107 pfu/mL以上,重组率92%以上,插入片段平均大于1.5 kb.肝脏cDNA文库的滴度在3.4× 107pfu/mL以上,重组率90%以上,插入片段平均大于1.4kb. 相似文献
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为了研究盐生植物耐盐基因表达调控,本实验以海水浇灌的海马齿植株为供试材料,构建了盐胁迫下的全长cDNA文库。构建方法如下：采用改良的CTAB法提取总RNA,SMART法反转录合成cDNA,LD-PCR方法合成双链cDNA。LD-PCR产物经蛋白酶K消化和SfiⅠ酶切后,经CHROMA SPIN＋TE-1000分离柱子除去小片段DNA后,回收0.5kb以上的片段,按照适当的比例连接λTripIEX2载体。连接产物利用MaxPlaxTMLambda Packaging Extracts进行体外包装,得到海马齿初级cDNA文库。初始文库的独立克隆数为2.4×106pfu,初级文库滴度大于4.80×106pfu/mL,重组率为93.75%,插入片段为0.5～5kb,扩增文库的滴度为1.21×109pfu/mL,所得文库质量较高。本研究表明该cDNA文库适合于盐生植物海马齿相关基因的克隆和分析。 相似文献
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摘要：了解肌肉组织和肝脏组织的基因类型，构建鸡肌肉组织和肝脏组织的功能基因表达谱在动物遗传育种中具有重要的意义。为比较基因组学的研究、功能基因组学的研究、QTLs定位等的动物分子育种做前期基础科研工作，研究以丝毛乌骨鸡(Gallus gallus )的胸肌和肝脏为实验材料，以Lambda ZAPⅡ为载体，构建了胸肌和肝脏的cDNA文库。同时，对这两个cDNA文库进行了噬菌体PCR鉴定和所提质粒的Eco RⅠ和XhoⅠ的双酶切鉴定。结果表明，胸肌和肝脏mRNA的OD260/OD280 分别为1.92和1.90。胸肌cDNA文库的滴度在3.6× 107 pfu/mL以上，重组率92%以上，插入片段平均大于1.5 kb。肝脏cDNA文库的滴度在3.4× 107 pfu/mL以上，重组率90%以上，插入片段平均大于1.4 kb。 相似文献
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华东葡萄白河-35-1株系抗霜霉病基因cDNA文库构建及EST分析 总被引：1，自引：0，他引：1
以中国原产抗霜霉病葡萄野生种华东葡萄（Vitis pseudoreticulata）白河-35-1为材料,用SMARTTM技术构建葡萄霜霉菌（Plasmopara viticola）诱导的葡萄叶片cDNA文库。经检测,原始文库滴度0.92×106 pfu/mL,重组率97%,插入片段500～2 200 bp,扩增文库滴度为5×109 pfu/mL。随机挑取克隆5'端测序,获得254条有效EST序列（GenBank登录号：FG269201~FG269449和FG396006~FG396010）。经BlastX分析,有56.3% ESTs与GenBank中功能已知基因有较高同源性,14.6%ESTs与功能尚未确定的假设蛋白或未知蛋白同源性很高,20.9%ESTs在GenBank中无匹配的同源产物。 相似文献
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松墨天牛（Monlchamus alternatus Hope）是林业重大害虫。本研究运用SMART技术构建了松墨天牛幼虫cDNA文库,库容量为1.09×10^6,文库滴度为5.45×10^6 pfu/mL,重组率为97.8%,平均插入片段大小约为1 000 bp,构建的文库质量较好。随机选取1 022个克隆进行测序,获得了1 012条表达序列标签（ESTs）,序列在GenBank中登录号为JZ143720～JZ144731。EST序列经过聚类拼接得到411条非重复单一序列,其中216条序列有功能注释。在216条有功能注释的序列中,有181条（83.80%）参与＂分子功能＂,122条（56.48%）参与＂细胞成分＂,168条参与＂分子生物学过程＂（77.78%）。研究结果为克隆和分析松墨天牛新的功能基因奠定了基础。 相似文献
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石门雄黄矿周边农田土壤重金属污染及健康风险评估 总被引：8，自引：2，他引：8
湖南石门雄黄矿区环境污染及风险是当前国家环保部门和地方政府极为关注的重要环境问题之一。本研究以该矿区周边农田表层土壤为研究对象,调查分析了矿区周边农田土壤重金属污染状况及其空间分布特征,并采用美国EPA土壤健康风险模型评估了其对人体的健康风险。土壤重金属污染评价结果显示:该区土壤受到中度的As污染和Cd污染,As平均含量为80.26 mg/kg,Cd平均含量为0.55 mg/kg;Cu、Zn、Pb含量均未超过国家《土壤环境质量》的二级标准;综合污染指数显示研究区域为中度污染。美国EPA模型评估结果显示:成人和儿童的日暴露量及非致癌健康风险主要途径均为经手–口摄入,Cd对成人和儿童均不存在非致癌风险和总风险,As对儿童非致癌总风险指数为3.36,造成严重的非致癌健康威胁;经呼吸暴露的致癌风险,Cd对儿童和成人均不造成致癌健康影响,As对成人和儿童的平均致癌风险指数为3.13×10~(-4)和5.58×10~(-4),均存在显著的致癌健康风险,且As、Cd对儿童的健康威胁均高于成人。可见,该矿区周边农田土壤中As、Cd污染及其风险应加强控制与治理。 相似文献
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水稻白叶枯病菌（Xanthomonas oryzae pv.oryzae,Xoo）引起的水稻白叶枯病是世界水稻生产中最严重的细菌性病害之一。前期研究发现Xoo PXO99A中一个编码Ⅲ型效应物的PXO_03420（hpaF）基因与病菌游动性、致病性和致敏性有关,为探讨hpaF基因编码产物在病菌与日本晴水稻中的互作关系,本研究采用Matchmaker酵母双杂交系统构建了日本晴水稻叶片高质量cDNA文库,文库重组率为87.6%,滴度为2.57×10^6pfu/mL。将文库与构建的Y187/pGBKT7_03420诱饵进行酵母双杂交,共得到26个阳性克隆。通过对阳性克隆测序所得结果进行结构域分析,初步鉴定HpaF与水稻叶片中具有WHY或PRK结构域的两个蛋白存在互作关系。本研究结果为进一步研究hpaF基因的功能奠定了基础。 相似文献
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Domínguez-Crespo Miguel A. Sánchez-Hernández Z. Erika Torres-Huerta Aidé M. Negrete-Rodríguez Ma. de la Luz X. Conde-Barajas Eloy Flores-Vela Abelardo 《Water, air, and soil pollution》2012,223(2):915-931
In order to enhance the removal of heavy metals such as Ni, Cu, Zn and Cd from wastewater, different cow dung/sewage sludge
ratios were tested to assess the effect of these metals on the adaptability of Eisenia fetida earthworms to the treatment process carried out in a typical plant located in Tamaulipas, Mexico. Two experimental water
treatment setups were proposed. The first set of experiments was planned to determine the adequate sewage sludge/cow dung
ratio(s), whereas the second arrangement was designed to evaluate the growth performance and fecundity of the earthworms under
high heavy metal concentrations. To achieve the objectives, the experiments were conducted for 90 days under controlled environmental
conditions. Maximum worm biomass and growth rates were attained in samples containing 25 wt.% of sewage sludge. Weight and
mortality of worms were significantly affected by the high levels of heavy metals, making difficult the metal accumulation
in the worm tissues. 相似文献
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Toxic effect of cadmium and nickel on soil enzymes and the influence of adding sewage sludge 总被引：4，自引：0，他引：4
Sewage sludge is increasingly used as an organic amendment to soil, especially to soil containing little organic matter. However, little is known about the utility of this organic amendment in the reclamation of soil polluted with heavy metals. We studied the effects of adding sewage sludge on enzymatic activities of a semi-arid soil contaminated with Cd or Ni in the laboratory. The activities of urease, phosphatase, β-glucosidase and protease-BAA were measured in soil containing concentrations of Cd or Ni in the range 0–8000 mg kg−1 soil, and their inhibition was compared with those of the enzymatic activities in the same soil amended with sewage sludge and containing similar concentrations of the heavy metals. The inhibition was tested for three different incubation times to determine changes in the effect of the heavy metals on hydrolase activity with the time elapsed after contamination. Ecological dose (ED) values of Cd and Ni were calculated from three mathematical models which described the inhibition of the enzymatic activities with increasing concentrations of heavy metal in the soil. For urease and phosphatase activities, the ED values for Cd and Ni increased after application of sewage sludge to soil, indicating a decrease in Cd and Ni toxicity. The other two enzymes (β-glucosidase and protease-BAA) were less sensitive to Cd or Ni contamination, and it was more difficult to determine whether addition of sewage sludge had affected the inhibition of these enzymes by the heavy metals. 相似文献