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Highly polymorphic microsatellite loci offer great promise for gene mapping studies, but fulfillment of this potential will require substantial improvements in methods for accurate and efficient genotyping. Here, we report a genotyping method based on fluorescently labeled PCR primers and size characterization of PCR products using an automated DNA fragment analyzer. We capitalize on the availability of three distinct fluorescent dyes to label uniquely loci that overlap in size, and this innovation increases by threefold the number of loci that can be analyzed simultaneously. We label size standards with a fourth dye and combine these with the microsatellite PCR products in each gel lane. Computer programs provide very rapid and accurate sizing of microsatellite alleles and efficient data management. In addition, fluorescence signals are linear over a much greater range of intensity than conventional autoradiography. This facilitates multiplexing of loci (since signal intensities often vary greatly) and helps distinguish major peaks from artifacts, thereby improving genotyping accuracy.  相似文献   
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以克山病病区粮配成基础饲料,另在基础饲料中分别补充Se或VE,或Se+VE喂养大鼠,在细胞及亚细胞水平上以Ca代谢为主研究并比较了Se和VE在克山病病因中的作用。测量了心肌细胞和心肌线粒体的Ca代谢及有关指标、心肌线粒体能量转换功能及心肌组织自由基含量。结果表明,在低Se病区粮中补充Se或VE均能在一定程度上预防病区粮中致病因素对心肌细胞及线粒体的损伤;并且补充Se或VE均能使心肌组织中自由基含量减少。提示Se和VE是通过清除体内过量自由基预防细胞和线粒体的损伤的。但值得注意的是,实验中所用病区粮VE含量不低于甚至高于非病区对照粮,在低Se情况下,所补VE的量需要相当大(如本实验中补充200μg/g)才能较明显地预防心肌细胞和心肌线粒体的损伤。通过对这些结果的分析,进一步肯定低Se是克山病形成的重要因素之一。  相似文献   
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Z F Wang  J Yang  Z Q Nie  M Wu 《Biochemistry》1991,30(4):1127-1131
A crude in vitro system which initiates chloroplast DNA synthesis near the D-loop site mapped by electron microscopy [Wu, M., Lou, J. K., Chang, D. Y., Chang, C. H., & Nie, Z. Q. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 6761-6765] consists of soluble proteins and proteins extracted from purified thylakoid membrane. In this paper, a DNA polymerase activity was purified to near homogeneity from the soluble protein fraction of this in vitro system by sequential chromatographic separations on heparin-agarose, DEAE-cellulose, and single-stranded DNA-agarose columns and sedimentation in a glycerol gradient. In the glycerol gradient, the enzyme activity sedimented at a position corresponding to a 110-kDa protein. Electrophoretic analysis of the highly purified fraction on SDS-polyacrylamide gel revealed a major polypeptide band with an apparent molecular mass of approximately 116 kDa. In situ DNA polymerase activity assay shows that the DNA polymerization function is associated with the 116-kDa band and an 80-kDa band which could be a subunit of the enzyme. Polymerization activity is inhibited by N-ethylmaleimide, ethidium bromide, and dideoxycytosine triphosphate and is relatively resistant to aphidicolin. Poly(dA).(dT)10 and gapped double-stranded DNA are preferred templates. The purified enzyme contains no exonuclease activity and can initiate DNA replication in a supercoiled plasmid DNA template containing the chloroplast DNA replication origin.  相似文献   
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互花米草入侵九段沙河口湿地对当地昆虫多样性的影响   总被引:15,自引:0,他引:15  
为认识因互花米草(Spartinaalterniflora)入侵而给九段沙河口湿地昆虫多样性带来的影响,我们于2004年5月到2005年10月间用网捕和收割植株两种方法对3个典型植物群落中昆虫多样性作了连续调查。研究期间,共采集到昆虫11,300头。经鉴定为97个种,隶属于12目69科。互花米草群落中昆虫的物种数、个体数量以及Shannon-Wiener多样性指数均显著低于土著植物芦苇(Phragmitesaustralis)群落和海三棱藨草(Scirpusmariqueter)群落中的;而Simpson优势度指数较土著植物群落中高。聚类分析结果表明:芦苇与海三棱藨草群落中昆虫群落结构更为相似。互花米草的入侵将可能导致九段沙湿地昆虫多样性的降低和群落结构的改变。  相似文献   
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藕莲有性繁殖栽培   总被引:2,自引:0,他引:2  
根据莲(Nelumbo nucifera)生物学特性,既可用藕作种进行无性繁殖,也可用种子播种进行有性繁殖。藕莲在我国有数千年的栽培历史,但素以无性系栽培。尚未见到有性繁殖栽培的报道。作者自1980年起进行了有性繁殖栽培试验。    相似文献   
9.
Mao JW  Wang LW  Jacob T  Sun XR  Li H  Zhu LY  Li P  Zhong P  Nie SH  Chen LX 《Cell research》2005,15(5):371-378
The transwell chamber migration assay and CCD digital camera imaging techniques were used to investigate the relationship between regulatory volume decrease (RVD) and cell migration in nasopharyngeal carcinoma cells (CNE-2Z cells). Both migrated and non-migrated CNE-2Z cells, when swollen by 47% hypotonic solution, exhibited RVD which was inhibited by extracellular application of chloride channel blockers adenosine 5‘-triphosphate (ATP), 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and tamoxifen. However, RVD rate in migrated CNE-2Z cells was bigger than that of non-migrated cells and the sensitivity of migrated cells to NPPB and tamoxifen was higher than that of nonmigrated cells. ATP, NPPB and tamoxifen also inhibited migration of CNE-2Z cells. The inhibition of migration was positively correlated to the blockage of RVD, with a correlation coefficient (r) = 0.99, suggesting a functional relationship between RVD and cell migration. We conclude that RVD is involved in cell migration and RVD may play an important role in migratory process in CNE-2Z cells.  相似文献   
10.
We looked for variations that could be associated with chicken egg number at 300 days of age (EN300) in seven genes of the hypothalamic-pituitary-gonadal axis, including gonadotrophin-releasing hormone-I (GnRH-I), GnRH receptor (GnRHR), neuropeptide Y (NPY), dopamine D2 receptor (DRD2), vasoactive intestinal polypeptide (VIP), VIP receptor-1 (VIPR-1), prolactin (PRL), and the QTL region between 87 and 105 cM of the Z chromosome. Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens, which were obtained from a company located in Guangdong Province of China. The C1704887T of VIPR-1 was found to have a highly significant association with EN300. The T5841629C of DRD2 and the C1715301T of VIPR-1 were significantly associated with EN300. A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300. H1H3 had the highest EN300. Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300. The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300. Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain GRB2-like 2 (SH3GL2) gene. We conclude that five SNPs, including C1704887T and C1715301T of VIPR-1, T5841629C of DRD2, and T32742468C and G32742603A of SH3GL2, would be useful as markers for breeding to increase chicken EN300.  相似文献   
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