[Objective] The aim was to provide molecular basis for the identification of species in the moss family Bryaceae by the construction of inter-simple sequence repeats （ISSR） fingerprinting. [Method] In order to seek standardizing PCR reaction set-up, an orthogonal design was used to optimize ISSR-PCR amplification system of Bryaceae in five factors （Mg2＋, dNTPs, primer, DNA template, Taq DNA polymerase） at four levels respectively. [Result] A suitable ISSR reaction system was obtained, namely： 20 μl reaction system containing 5 ng of DNA template, 0.2 μmol/L primer, 2.25 mmol/L MgCl2, 0.6 U of Taq DNA polymerase, 0.4 mmol/L dNTPs. Proper annealing temperature was found at 48-50 ℃.The above system and six ISSR-PCR primers were used for the PCR amplification of 14 samples from Bryaceae and the related species in Mniaceae. A total of 86 bands were amplified, all showed polymorphism. NJ cluster analysis showed a star-shaped cladogram. [Conclusion] The results manifested that ISSR fingerprinting could provide the appropriate degree of polymorphism at low taxonomic level, so it would be a useful tool to provide additional evidence for resolving taxonomic relationships at the species level of Bryaceae.
选用未离乳的SD大鼠十二指肠,采用2次酶消化及不同孔径滤网分级过滤等方法分离十二指肠微血管,通过体外培养、纯化成功获得了高纯度的原代培养的小肠微血管内皮细胞,并应用Western-blot技术,首次检测到了小肠微血管内皮细胞具有铁转运相关蛋白DMT1(divalent metal transporter1),FPN1(feerroportin1)和Tfr1的表达.这一新发现极大地丰富了小肠铁吸收及小肠铁吸收调节机制的理论,对进一步开展小肠铁代谢及其调节机制和铁代谢疾病的研究具有重要的理论和应用价值.
