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1.
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   

2.
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   

3.
采用人工土壤法,研究重金属镉(Cd2+)对成熟赤子爱胜蚓(Eisenia fetida)的急性毒性,表明蚯蚓对镉具有较强的耐受性。Trizol法提取蚯蚓RNA后,采用Clontech公司的CreatorTM SMARTTM cDNA文库构建试剂盒,成功构建了低剂量Cd2+(200 mg•kg-1)诱导的蚯蚓cDNA文库,其库容量为3.02×105 pfu/mL,扩增后滴度为8.67×109 pfu/mL,重组率为97.3%, 绝大多数的cDNA插入片段≥0•5 kb,平均长度≈1•0 kb。所够建的cDNA质粒文库容量及插入片段大小符合合格文库要求, 为进一步筛选蚯蚓体内解毒、免疫相关基因,寻找生物标志物奠定了基础。  相似文献   

4.
采用SMART技术首次构建了高质量的紫茎泽兰cDNA文库。经过涂平板测定和酶切反应鉴定表明,原始文库的滴度为1.23×107 cfu/mL,重组率达98 %,插入片段平均大小在900 bp左右,采用涂平板均匀扩增得到的扩增文库滴度达到了1×1012,覆盖率达99 %以上。利用该文库筛选得到了紫茎泽兰核酮糖二磷酸羧化酶(RuBP羧化酶)cDNA片段。  相似文献   

5.
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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环境胁迫诱导的花生全长cDNA文库的构建及序列分析   总被引:1,自引:0,他引:1  
以花生新品种闽花6号为材料,用生物和非生物因素胁迫处理植株,采用SMART技术构建了花生受低温、干旱、各种激素、缺钙及黄曲霉等胁迫诱导的混合全长cDNA文库,原始文库滴度为8.86×106 cfu,重组率达97.6%,插入片段多在1~2kb之间,平均大小为1 300bp.该cDNA文库的构建可用于花生抗逆基因的发掘,为揭示花生抗逆分子机理及花生品种改良奠定基础.  相似文献   

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欧李叶片全长cDNA文库的构建和部分克隆的序列分析   总被引:4,自引:1,他引:3  
采用SMART技术,构建了欧李(Prunus humilis Bunge)品系T8-1叶片全长cDNA文库,原始文库滴度达6.17×10~7 pfu/mL,库容达3.7×10~7,重组率达95%,平均插入片段大小约为1.2 kb.随机挑取400个单克隆进行5'端测序,共获得364个欧李ESTs.绝大部分ESTs的长度在500~1300 bp之间,平均长度为877 bp,其中具有完整ORF结构的序列有221条,占60.71%.通过与NCBI等非冗余核酸数据库和蛋白质数据库进行比对、查询和注释,获得已知功能基因或具推测功能的基因181个(339个ESTs),相似性较低的未鉴定基因5个(9个ESTs),新基因14个(16个ESTs).  相似文献   

11.
平原地区河道堤防滩地的水土流失,直接淤积河床,影响行洪安全。堤防滩地的水土流失是自然因素和人为因素共同作用的结果,以新修堤防的水土流失最为严重,对其防治须实行工程措施、植物措施和人为预防相结合。  相似文献   

12.
重庆地处长江上游、三峡库区,由于三峡库区水土流失严重、生态环境脆弱、自然灾害频繁,且三峡库区的水土保持和生态建设事关全市经济社会发展全局、事关三峡工程建成后的安全运行和库区的长治久安,因此是我市水土保持生态建设的重点。虽然,“长治”工程的有效实施,使我市的水土保持生态建设取得了较大的成绩,但离全面建设小康社会的要求尚有不少的差距,因此,我们要紧紧抓住国家实施可持续发展战略和西部大开发战略的历史机遇,进一步增强紧迫感、责任感和使命感,调动全社会各界力量,防治水土流失,改善生态环境,建设巴渝秀美山川。  相似文献   

13.
论水土、水土生态与水土生态保持   总被引:4,自引:7,他引:4  
在论述水土在陆地生态系统中的地位和作用的基础上,提出了水土生态的概念,认为植被与水土不可分割的整体观念是水土生态的重要特征。同时,对水土生态保持的含义作了新的定义,并将水土生态保持划分为四大类型,即生态型、自然型、生产型、建设型。从水土生态的高度,从源头上、要素的联系中去认识和防治水土流失,是一种主动的、有机的、整体的水土保持观念,是水土保持认识观的深化和发展,将使水土生态保持事业进入一个崭新的时代。  相似文献   

14.
The charge characteristics of A1 or Ap and B2 horizon samples of total 23 Ultisols, Alfisols and Oxisols in Korea and Thailand were studied by measuring the retention of NH4+ and NO3? at different pH values (4–8) and NH4NO3 concentrations (0.1–0.005 m ). The magnitude of their negative charge (σ?; meq/100g) was dependent on pH and NH4NO3 concentration (C; m ) as represented by a regression equation: log σ?=apH +blogC +c. The values of the coefficient a (0.04–0.226), b (0.03–0.264) and c (–0.676–1.262) were correlated with the kinds of the soil and horizon and with the region where the soil exists. The retention of NO3? was less than 1 and 2–3 meq/100 g for the A1 or Ap and B2 horizon samples, respectively. The sum of exchangeable base and Al (‘effective’ CEC) was close to and higher than the magnitude of permanent charge (=σ? measured at pH = 4.3 and at C = 0.005 m ) for one-third and two-thirds of samples, respectively. A σ? value of 16 meq/100 g clay at pH = 7 and C = 0.01 m was found appropriate to separate the B2 horizons of Thai Ultisols and Oxisols from those of Korean Ultisols and Alfisols. Korean Alfisols and Ultisols and Thai Ultisols were distinguished from each other on the status of exchangeable base and Al  相似文献   

15.
水土保持法律规范和相关行政法律规范,共同构成了水土保持监督管理法律体系,其内容庞杂,相互交叉、融合,程序复杂。通过一组网络图及其简要注释,直观地表述了水土保持监督管理工作的相关程序、时效、逻辑关系和法律依据;通过对网络图呈现的逻辑问题所反映出的法律缺陷进行分析,提出应当按照立法法的有关规定,对现行法律特别是水土保持法律法规进行必要的修改,以适应不断发展的水土保持监督管理工作需要的建议。  相似文献   

16.
简析了婺源县城镇开发区建设过程中造成水土流失的状况、危害、原因后,提出了其防治措施,并阐明开展城市(镇)水土保持的紧迫性。  相似文献   

17.
我国滑坡、崩塌的区域特征、成因分析及其防御   总被引:2,自引:0,他引:2  
论述了我国滑坡、崩塌的区域分布特征,滑坡和崩塌的危害程度,滑坡和崩塌类型和成因分析,并且提出了灾害的防御措施,以期达到环境保护成为社会发展过程中的一个重要组成部分。  相似文献   

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Soils were collected from 2-year (2-y) and 3-year (3-y) old red-pine seedling plots in two tree nurseries, Hayward in the north and Wilson in the southwestern part of Wisconsin State respectively, and equilibrated with 0.01 M Ca(NO3)2 for soil solution Zn and Mn (solu-Zn and Mn), and with 0.01 M Ca(NO3)2+0.005 M EDTA for soil adsorbed Zn and Mn (ad-Zn and Mn). Buffering capacity of soil Zn and Mn (b-Zn and Mn) was obtained from the ratio of ad-Zn and Mn to the solu-Zn and Mn. The concerned traces in pine seedling needles (ndls), stems(sts) and roots (rts) were simultaneously measured.  相似文献   

20.
Abstract

Lucerne (Medicago sativa L.) and phalaris (Phalaris aquatica L.) were grown separately and together in a pot trial on a yellow‐brown pumice soil with three rates of sodium (Na) and two rates of potassium (K) in factorial combination. Lucerne alone had a low Na concentration but growing phalaris as a companion grass produced herbage with a Na concentration adequate for stock. Na application increased the Na concentration in phalaris and the mixture of phalaris and lucerne much more than in lucerne alone; had little effect on K concentration; slightly reduced Mg concentration; and greatly reduced Ca concentration but not as much in lucerne as in phalaris or the mixture with lucerne. K application increased K concentration and reduced Na, Ca and Mg concentrations throughout. Yield of phalaris grown alone and in combination with lucerne was increased significantly by Na application when K concentration in the plants was low. Yield of lucerne was not affected by Na application and it is concluded that Na did not substitute for K in this species. It is concluded that field trials are warranted to investigate the possibility of growing a special purpose mixture of lucerne and phalaris on New Zealand yellow‐brown pumice soils to provide feed that has adequate Na for grazing animals.  相似文献   

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