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221.
Maureen P. Martin Anita Harding Robert Chadwick Mel Kronick Michael Cullen Ling Lin Emmanuel Mignot M. Carrington 《Immunogenetics》1997,47(2):131-138
The human genome contains a large number of interspersed microsatellite repeats which exhibit a high degree of polymorphism
and are inherited in a Mendelian fashion, making them extremely useful genetic markers. Several microsatellites have been
described in the HLA region, but allele nomenclature, a set of broadly distributed controls, and typing methods have not been standardized, which
has resulted in discrepant microsatellite data between laboratories. In this report we present a detailed protocol for genotyping
microsatellites using a semi-automated fluorescence-based method. Twelve microsatellites within or near the major histocompatibility
complex (MHC) were typed in the 10th International Histocompatibility Workshop homozygous typing cell lines (HTCs) and alleles
were designated based on size. All loci were sequenced in two HTCs providing some information on the level of complexity of
the repeat sequence. A comparison of allele size obtained by genotyping versus that obtained by direct sequencing showed minor
discrepancies in some cases, but these were not unexpected given the technical differences in the methodologies. Fluorescence-based
typing of microsatellites in the MHC described herein is highly efficient, accurate, and reproducible, and will allow comparison
of results between laboratories.
Received: 10 May 1997 / Revised: 1 August 1997 相似文献
222.
不同降温速率对脐血干细胞冷冻复苏后生物学特性的影响 总被引:4,自引:0,他引:4
考察了不同降温速率对脐血造血干细胞各种生物学特性的影响。在4℃~-40℃的降温范围内,分别选择-0.5℃/min, -1℃/min, -5℃/min的降温速率进行降温,对复苏后的脐血单个核细胞的回收率、活性和CD34+含量的变化以及BFU-E、CFUGM和CFU-MK集落的回收率进行了考察,发现在-1℃/min的降温速率下,脐血MNC回收率可达93.3%±1.8%,活性可达95.0%±3.9%, CD34+细胞回收率达80.0%±17.9%,BFUE回收率为87.1%±5.5%,CFUGM回收率达88.5%±8.9%,CFUMK的回收率也达到86.2%±7.4%。并且对复苏后的细胞进一步进行体外培养,发现在-1℃/min的降温速率下复苏的细胞仍然具有与未经冷冻细胞相似的扩增能力,而-0.5℃/min和-5℃/min这两种降温速率条件下复苏的细胞与未经冷冻的细胞相比差距较大。因而-1℃/min的降温速率对冻存脐血干细胞比较合适。 相似文献
223.
Associations between δ13C values and leaf gas exchanges and tree-ring or needle growth, used in ecophysiological compositions, can be complex depending
on the relative timing of CO2 uptake and subsequent redistribution and allocation of carbon to needle and stem components. For palaeoenvironmental and
dendroecological studies it is often interpreted in terms of a simple model of δ13C fractionation in C3 plants. However, in spite of potential complicating factors, few studies have actually examined these relationships in mature
trees over inter- and intra-annual time-scales. Here, we present results from a 4 years study that investigated the links
between variations in leaf gas-exchange properties, growth, and dated δ13C values along the needles and across tree rings of Aleppo pine trees growing in a semi-arid region under natural conditions
or with supplemental summer irrigation. Sub-sections of tissue across annual rings and along needles, for which time of formation
was resolved from growth rate analyses, showed rapid growth and δ13C responses to changing environmental conditions. Seasonal cycles of growth and δ13C (up to ~4‰) significantly correlated (P<0.01) with photosynthetically active radiation, vapour pressure deficit, air temperature, and soil water content. The irrigation
significantly increased leaf net assimilation, stomatal conductance and needle and tree-ring growth rate, and markedly decreased
needle and tree-ring δ13C values and its sensitivity to environmental parameters. The δ13C estimates derived from gas-exchange parameters, and weighted by assimilation, compared closely with seasonal and inter-annual
δ13C values of needle- and tree-ring tissue. Higher stomatal conductances of the irrigated trees (0.22 vs. 0.08 mol m−2 s−1 on average) corresponded with ~2.0‰ lower average δ13C values, both measured and derived. Derived and measured δ13C values also indicated that needle growth, which occurs throughout the stressful summer was supported by carbon from concurrent,
low rate assimilation. For Aleppo pine under semi-arid and irrigated conditions, the δ13C of tree-ring and needle material proved, in general, to be a reasonable indicator of integrated leaf gas-exchange properties. 相似文献
224.
利用27对SSR分子标记对新疆4个野杏群体遗传多样性和遗传结构进行分析,评价新疆野杏遗传多样性水平和分化程度,为新疆野杏合理保护与利用提供科学依据。结果显示:(1)27对SSR引物共检测到431个等位基因(Na),各位点平均等位基因数(Na)和多态性信息含量(PIC)分别为15.96和0.84;物种水平上Shannons信息指数(I)和期望杂合度(He)分别为2.21和0.78。(2)群体水平上等位基因数(Na)、有效等位基因(Ne)、Shannons信息指数(I)、期望杂合度(He)和观察杂合度(Ho)分别为10.98、5.85、1.92、0.79和0.55;其中新源县野杏群体遗传多样性最丰富,巩留县群体遗传多样性最低。(3)基于F统计量分析的遗传分化系数(Fst)为0.05,基因流(Nm)为5.26;分子方差分析显示新疆野杏群体大部分遗传变异来自群体内(95.4%),群体间的遗传变异仅占4.6%。(4)新疆野杏群体遗传距离为0.06~0.49,平均为0.24;遗传相似度为0.61~0.94,平均为0.80;遗传相似度的聚类分析和遗传距离的主坐标分析结果一致,均将供试4个群体划分为两组;Mantel检测显示,新疆野杏群体遗传距离与地理距离无显著相关(r=0.332,P=0.16)。研究表明,新疆野杏资源具有丰富的遗传多样性,群体遗传分化程度较低,群体间遗传距离较小,这与新疆野杏群体的大小和悠久的演化历史以及群体间频繁的基因交流相关。 相似文献
225.
Biology Bulletin - CRISPR is an adaptive immune defense system found in bacteria and archaea that is resistant to heterologous invasive genetic material. Later studies showed that the CRISPR system... 相似文献
226.
Promoter methylation of the O6-methylguanine-DNA-methyltransferase (MGMT) gene has been considered a prognostic marker and has become more important in the treatment of glioblastoma. However, reports on the correlation between MGMT and clinical outcomes in Chinese glioblastoma patients are very scarce. In this study, quantitative methylation data were obtained by the pyrosequencing of tumor tissues from 128 GBM patients. The median overall survival (OS) was 13.1 months, with a 1-year survival of 45.3%. The pyrosequencing data were reproducible based on archived samples yielding data for all glioblastomas. MGMT promoter methylation was detected in 75/128 cases (58.6%), whereas 53/128 (41.4%) cases were unmethylated. Further survival analysis also revealed that methylation was an independent prognostic factor associated with prolonged OS but not with progression-free survival (PFS) (p = 0.029 and p = 0.112, respectively); the hazard radios were 0.63 (95% CI: 0.42–0.96) and 0.72 (95% CI: 0.48–1.09), respectively. These data indicated that MGMT methylation has prognostic significance in patients with newly diagnosed high-grade glioblastoma undergoing alkylating agent-based chemotherapy after surgical resection. 相似文献
227.
Receptor-mediated endocytosis begins with the binding of ligand to receptors in clathrin-coated pits followed by the budding of the pits away from the membrane. We have successfully reconstituted this sequence in vitro. Highly purified plasma membranes labeled with gold were obtained by incubating cells in the presence of anti-LDL receptor IgG-gold at 4 degrees C, attaching the labeled cells to a poly-L-lysine-coated substratum at 4 degrees C and then gently sonicating them to remove everything except the adherent membrane. Initially the gold label was clustered over flat, clathrin-coated pits. After these membranes were warmed to 37 degrees C for 5-10 min in the presence of buffer that contained cytosol extract, Ca2+, and ATP, the coated pits rounded up and budded from the membrane, leaving behind a membrane that was devoid of LDL gold. Simultaneous with the loss of the ligand, the clathrin triskelion and the AP-2 subunits of the coated pit were also lost. These results suggest that the budding of a coated pit to form a coated vesicle occurs in two steps: (a) the spontaneous rounding of the flat lattice into a highly invaginated coated pit at 37 degrees C; (b) the ATP, 150 microM Ca2+, and cytosolic factors(s) dependent fusion of the adjoining membrane segments at the neck of the invaginated pit. 相似文献
228.
玉米花粉单倍体植株染色体上异染色质的变异 总被引:3,自引:1,他引:3
我们用Giemsa BSG C-带技术检查了玉米花药培养获得的花粉单倍体植株根尖细胞染色体上异染色质的变异,观察结果表明,有的植株所显示的C-带数目是与供体植株的相一致,有的植株所显示的C-带数目则发生了显著变化,其中有的增加,有的减少。并讨论了异染色质发生变异的可能原因。还相应地观察到间期核中染色中心的变化是与中期染色体上C-带数目的变化相一致。 相似文献
229.
Background
Due to shared transmission routes, hepatitis C virus (HCV) infection is highly prevalent among people infected with human immunodeficiency virus (HIV). Highly active antiretroviral therapy (HAART) is associated with hepatotoxicity, leading to the negative effects on patients with HIV/HCV co-infection. In order to provide valuable information for HCV management in this particular population, we investigated the HCV genotypes in HIV-infected individuals from Henan and Guangxi, the two provinces with the most HIV-infected cases in China.Methods
Individuals, who acquired HIV infection through various risk routes, were recruited from Henan and Guangxi. Test of antibodies against HCV (anti-HCV) was conducted, and detection of HCV RNA was performed by PCR amplification. HCV subtypes were determined by direct sequencing of amplicons, followed by phylogenetic analysis.Results
We recruited a total of 1,112 HIV-infected people in this present study. Anti-HCV was detected from 218 (50.1%) patients from Henan and 81 (12.0%) patients from Guangxi, respectively. The highest prevalence of HIV/HCV co-infection was observed from FBDs (former blood donors) (87.2%) in Henan and IDUs (intravenous drug users) (81.8%) in Guangxi, respectively. The seroprevalence rate of HCV among people with sexual contact was significantly higher in Henan than in Guangxi (18.7% vs. 3.5%, P<0.05). The positive rate of HCV RNA in Henan and Guangxi was 30.6% (133/435) and 11.2% (76/677), respectively. Moreover, we found that 20 anti-HCV negative samples were HCV positive by PCR amplification. HCV subtype 1b (52.7%) was predominant in Henan, followed by subtype 2a (41.9%). The most frequently detected subtypes in Guangxi were 6a (35.6%) and 3b (32.9%).Conclusion
The HCV genotype distributions were different in HIV-infected people from Henan and Guangxi. HIV/HCV co-infection was not only linked to the transmission routes, but also associated with the geographic position. 相似文献230.