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重叠区扩增法合成抗菌肽B基因 总被引:1,自引:0,他引:1
人工设计并合成了抗菌肽 B基因的 4个寡聚核苷酸片段 ,通过重叠区扩增法 ,扩增出了相当于抗菌肽基因全长的寡聚核苷酸片段。经克隆测序 ,证明成功地实现了抗菌肽基因的人工合成、拼接和克隆。 相似文献
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Andrea Arias Aguirre Bruno Studer Javier Do Canto Ursula Frei Thomas Lübberstedt 《Plant Breeding》2014,133(6):765-770
Perennial ryegrass (Lolium perenne L.) displays a two‐locus gametophytic self‐incompatibility (SI) system that remains intact at the tetraploid level. Two models are plausible for SI in autotetraploids. In Model I: both alleles at the S locus and both at the Z locus in diploid pollen matching the female genotype results in incompatibility. In Model II: only one allele at S and one at Z locus in diploid pollen matching the female results in incompatibility. The goals were to determine which of the models best explains SI in our autotetraploid ryegrass population and to evaluate the efficiency of high‐resolution melting (HRM) genotyping for discriminating different iso‐allelic genotypes. The progeny of a cross between two autotetraploids was characterized with three HRM‐based markers co‐segregating with Z. Segregation ratios were used to make inferences about the mode of action of the SI system. The observed segregation differed significantly (P < 0.001) from the expected under Model I, but not from the expected under Model II (P = 0.463). Thus, Model II explains SI in this population, and HRM is an efficient tool to distinguish different iso‐allelic genotypic classes. 相似文献
158.
乐果液相色谱与气相色谱检测方法的比较 总被引:2,自引:0,他引:2
摘 要:【研究目的】通过应用液相色谱法(HPLC)和气相色谱法(GC)对乐果测定的参数进行比较,为乐果检测方法的选择提供参考。【方法】分别应用气相色谱和高效液相色谱法对乐果检测条件进行优化,对这两种测定方法的灵敏度、线性相关性、稳定性及添加回收率的等参数测定结果进行比较。【结果】用液相色谱法测定乐果最低检测限为0.01mg/L,比用气相色谱法(最低检测到0.1mg/L)测定时有更高的灵敏度;HPLC检测(r=0.9990)较GC(r=0.9937)有更好的线形相关性;在乐果浓度低于0.1mg/L时,用液相色谱法对乐果进行测定有很好的稳定性,在浓度高于0.1mg/L,两种测定方法的稳定性相当。【结论】用液相色谱法测定乐果完全可以达到农药登记残留试验准则的要求,它比气相色谱法更适合乐果的微量及常量的检测。 相似文献
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根据GeneBank(U37518)中TRAIL(肿瘤坏死因子相关的凋亡诱导配体)的cDNA序列,设计扩增引物。采用RT-PCR从人胎盘中扩增出TRAIL基因的全长cDNA。产物纯化后连至pGEM-TEasy载体,转化大肠杆菌DH5α。筛选阳性克隆菌,酶切鉴定并进行序列测定。结果表明,本试验成功地克隆了TRAIL基因的1039bp全长cDNA。 相似文献
160.
Involvement of a vascular hypersensitive response in quantitative resistance to Ralstonia solanacearum on tomato rootstock cultivar LS‐89
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K. Nakaho S. Seo K. Ookawa Y. Inoue S. Ando Y. Kanayama S. Miyashita H. Takahashi 《Plant pathology》2017,66(1):150-158
Ralstonia solanacearum causes bacterial wilt disease in Solanaceae spp. Expression of the Phytophthora inhibitor protease 1 (PIP1) gene, which encodes a papain‐like extracellular cysteine protease, is induced in R. solanacearum‐inoculated stem tissues of quantitatively resistant tomato cultivar LS‐89, but not in susceptible cultivar Ponderosa. Phytophthora inhibitor protease 1 is closely related to Rcr3, which is required for the Cf‐2‐mediated hypersensitive response (HR) to the leaf mould fungus Cladosporium fulvum and manifestation of HR cell death. However, up‐regulation of PIP1 in R. solanacearum‐inoculated LS‐89 stems was not accompanied by visible HR cell death. Nevertheless, upon electron microscopic examination of inoculated stem tissues of resistant cultivar LS‐89, several aggregated materials associated with HR cell death were observed in xylem parenchyma and pith cells surrounding xylem vessels. In addition, the accumulation of electron‐dense substances was observed within the xylem vessel lumen of inoculated stems. Moreover, when the leaves of LS‐89 or Ponderosa were infiltrated with 106 cells mL?1 R. solanacearum, cell death appeared in LS‐89 at 18 and 24 h after infiltration. The proliferation of bacteria in the infiltrated leaf tissues of LS‐89 was suppressed to approximately 10–30% of that in Ponderosa, and expression of the defence‐related gene PR‐2 and HR marker gene hsr203J was induced in the infiltrated tissues. These results indicated that the response of LS‐89 is a true HR, and induction of vascular HR in xylem parenchyma and pith cells surrounding xylem vessels seems to be associated with quantitative resistance of LS‐89 to R. solanacearum. 相似文献