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The Qinghai-Tibet Plateau is a more sensitive area of climate change and fragile zone of ecological environment. Qinghai Province is an important component of the Qinghai-Tibet Plateau, and the study of streamflow in the province is of great significance for understanding the situation of water resources. Based on hydrological data of hydrological stations in Qinghai, the spatiotemporal variations of streamflow from 1956 to 2013 were analyzed by Mann-Kendall analysis and Morlet wavelet analysis. The results indicated that the annual streamflow of the Yangtze River basin, Qaidam basin, Qinghai Lake basin, and Lantsang River basin showed significantly increasing trends during 1956-2013, while the Yellow River basin showed no significant trends or downward trends. Changes in precipitation, glacier melt, and snowmelt caused by climate warming might be a major driver of changes of streamflow in the source regions of the Yangtze River and Lantsang River basins. The trend variations of flow in the Yellow River basin may be affected by precipitation, evaporation caused by increased temperature, and anthropogenic impacts. The abrupt changes of streamflow at Tuotuohe, Delingha, and Nachitai stations were mainly in the early 2000s, while the abrupt changes of streamflow in Zhimenda station, stations in the Yellow River basin, Qinghai Lake basin, and Lantsang River basin appeared primarily in the early 1960s and the early 2000s. Most of the streamflow in Qinghai Province was abundant after about 2007, and the dominant periods with low frequency fluctuation (about 12 years) and inter-decadal (about 44 years) oscillations in Zhimenda and Tangnaihai stations and low frequency fluctuation (about five years) in other stations.
The open reading frame of chicken calpastatin (CAST) gene composed of 2,301 base pairs was ligated into a prokaryotic expression vector pET21a (+) to yield pET21a - CAST. The C-terminal His-tagged CAST protein was then expressed in E. coli. BL21 (DE3). SDS-PAGE analysis confirmed the successful expression of the fusion protein following induction with isopropyl-β-Dthiogalactopyranoside (IPTG). The recombinant protein consisted of 776 amino acid residues with an apparent molecular weight of approximately 110 kDa. It was primarily expressed as a soluble protein with a heat-stable feature. After being purified by Ni2+-NTA affinity resin, a polyclonal antibody was raised against the purified His-tagged CAST protein in rabbits. The reactivity and specificity of the polyclonal antibody were both subsequently characterized by ELISA. The study provides an important experimental tool for further research on the quantification of chicken CAST protein.
High-resolution melting analysis (HRMA) was used to screen potential SNPs in the exons of chicken CAPN1 (μ-calpain/large subunit) gene. A total of 312 DNA samples from Beijing-you chickens were used for detection. Twelve pairs of primer were designed to amplify twelve different exons and SNPs were detected in five of them. HRMA was also compared with PCR-SSCP analysis for genotyping of a known SNP site in the chicken adipocyte fatty acid binding protein gene (A-FABP). Amplicons of 275-bp fragment, bracketing the polymorphic site, were grouped by PCR-SSCP into three genotypem designated as CC, TT and CT. Small amplicons (56 bp) within the 275-bp fragments were designed to maximize the Tm difference between homozygotes and to genotype all possible three genotypes after a single melting analysis successfully. Results from different methods were cross-validated and sequencing results from randomly selected heterozygotes and homozygotes confirmed the specificity of HRM technique. The full consistency proved that HRMA was a useful tool for rapid, close-tube genotyping of polymorphic sites. It has great potential for SNPs detection and scanning especially on a large scale.
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