藏山羊KLF8基因克隆及组织表达分析

本试验旨在获得藏山羊KLF8基因序列,并分析其生物学特征,同时阐明该基因在不同组织中的表达情况。利用RT-PCR技术克隆藏山羊KLF8基因序列,利用实时荧光定量PCR (quantitative real-time PCR,qPCR)检测其在藏山羊各个组织中的表达丰度。结果表明,获得藏山羊KLF8基因序列1 069 bp,其中包含CDS区1 008 bp,5'UTR序列28 bp和3'UTR序列33 bp,共编码335个氨基酸,为不稳定亲水碱性蛋白。KLF8基因在藏山羊的肺脏组织中表达水平最高,极显著高于其他组织(p<0.01)。本研究为进一步阐明KLF8基因在藏山羊中的生物学功能提供了依据。     

青藏高原气候变化对农牧民开垦的影响

自1980年代后期以来,青藏高原持续变暖趋势明显,且增温幅度高于我国其他地区。基于遥感数据和种植适宜性模型的一些研究表明,该地区足够的农业热量资源增加了适宜耕种土地的面积,为农牧民的开垦行为创造了有利的条件。然而,对农牧民是否开垦以及开垦原因的实证研究,以及农牧民如何平衡气候变化带来的开垦机遇、风险和人口压力,仍然缺少认识。以青藏高原的3个典型产粮区域("一江两河"地区、河湟谷地、壤塘县)为研究区,利用605户农户家庭调研数据,分析了影响农牧民开垦行为的影响因素。计量结果表明,农牧民感知冬季持续时间的整体变化趋势、家庭总人口、抚养比与农牧民开垦行为呈显著正相关关系;农牧民感知降水的整体变化趋势、人均打工年收入、户主受教育水平、是否借贷、居住地距集镇距离与农牧民开垦行为呈显著负相关关系。随着城镇化的不断加快,农牧民对非农工作的认可度越来越高,加之政府的管制,使得开垦现象并不普遍。由于农牧民的开垦对青藏高原的生态环境极有可能造成不可逆的负面影响,应该提供更多的非农就业机会,促进农业集约化,加强监管,以降低开垦的可能性。     

北极狐β-防御素103基因 (CBD103) 启动子活性及转录调控元件分析

本研究旨在筛选调控北极狐毛色基因CBD103的启动子活性区域及转录因子结合位点,为揭示CBD103基因调控北极狐毛色形成的分子遗传机制提供依据。克隆获得了北极狐CBD103基因5′侧翼区2 123 bp的片段,并构建了4个不同长度的启动子缺失片段表达载体,通过双荧光素酶检测系统对启动子活性进行检测。对启动子活性最高区域预测出的3个特异性蛋白1 (Sp1)转录因子结合位点分别进行点突变并构建3个突变载体,利用双荧光素酶检测系统测定其活性。结果显示,在构建的4个不同长度启动子缺失片段中1 656 (-1 604/+51)区域活性最高,在此区域构建的3个突变载体的启动子活性较野生型(片段1 656)均显著降低,说明-1 604/+51区域为北极狐CBD103基因的核心启动子区,-1 552/-1 564、-1 439/-1 454和-329/-339区域为正调控区域。文中成功获得了北极狐CBD103基因的核心启动子区域和正调控区域,这为进一步研究该基因调控北极狐被毛颜色分子遗传机制奠定了基础。     

Generalized spatial mark–resight models with incomplete identification: An application to red fox density estimates

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Taenia crassiceps organic acids detected in cysticerci

Taenia crassiceps cysticerci is used as an experimental model to cysticercosis studies; however there are subcutaneous cases of cysticercosis caused by these cysticerci. It remains unclear in the literature the energetic and fatty acid metabolism in cestodes. Its metabolic study may provide knowledge of pathways that may serve as potential anti-helminthic drugs sites of action. In this work we studied the citric acid cycle organic acids and the fatty acid oxidation in cysticerci removed from mice with 21 and 42 days of infection in two different evolutive stages: growing and final. The organic acids were extracted using perchloric acid and analyzed by HPLC methodology. We found significant statistically differences in oxalate, malate, lactate, and beta-hydroxybutirate concentrations between cysticerci. These results indicate the aerobic metabolism in vivo in spite of the low oxygen concentration of its habitat, and also indicate the presence of fatty acid oxidation as an alternative energetic source.     

利用SSR标记分析云南、西藏和新疆小麦的遗传多样性

用185对SSR引物对52份中国西部特有小麦的遗传多样性进行了研究分析。在31份云南小麦材料中,共检测到488个等位变异,每一个SSR引物可检测到1至9个等位变异,平均为2.64个;平均PIC值为0.2764。在15份西藏小麦材料中,共检测到472个等位变异,每个引物可扩增出1到8个等位变异,平均为2.55个;平均PIC值为0.3082。在6份新疆小麦材料中,共检测到308个等位变异,每一个SSR引物可检测1到5个等位变异,平均为1.66个;平均PIC值为0.1944。185对SSR引物在云南、西藏和新疆小麦的21条染色体、7个部分同源群和3个染色体组上检测到的等位位点的多态性存在明显差异。云南、西藏和新疆小麦均以3B染色体较高,而1D染色体最低;在7个部分同源群中,均以第三部分同源群最高,第六部分同源群最低;在A、B和D染色体组上,均以B染色体组最高,D染色体组最低,A染色体组居中。利用185对SSR引物计算了云南、西藏和新疆小麦群体内及其群体间的遗传距离(GD)和平均遗传距离,结果显示,西藏小麦和云南小麦群体内的平均遗传距离要高于新疆小麦,而云南小麦和西藏小麦间的平均遗传距离低于两者与新疆小麦的平均遗传距离。聚类分析结果也表明,云南小麦和西藏小麦的亲缘关系较近,但两者与新疆小麦的亲缘关系相对较远。     

Characterization of <Emphasis Type="Italic">target</Emphasis> nuclear DNA from faeces reduces technical issues associated with the assumptions of low-quality and quantity template

Faecal material has increasingly become an important non-invasive source of DNA for wildlife population genetics. However, DNA from faecal sources can have issues associated with quantity (low-template and/or low target-to-total DNA ratio) and quality (degradation and/or low DNA-to-inhibitor ratio). A number of studies utilizing faecal material assume and compensate for the above properties with minimal characterization of quantity or quality of target DNA, which can unnecessarily increase the risk of downstream technical problems. Here, we present a protocol which quantifies faecal DNA using a two step approach: (1) estimating total DNA concentration using a Picogreen^TM fluorescence assay and (2) estimating target nuclear DNA concentration by comparing amplification products of field samples at suspected concentrations to those of control DNA at known concentrations. We applied this protocol to faecal material collected in the field from two species: woodland caribou (Rangifer tarandus) and swift fox (Vulpes velox). Total DNA estimates ranged from 6.5 ng/μl to 28.6 ng/μl (X = 16.2 ng/μl) for the caribou extracts and 1.0–26.1 ng/μl (X = 7.5 ng/μl) for the swift fox extracts. Our results showed high concordance between total and target DNA estimates from woodland caribou faecal extracts, with only 10% of the samples showing relatively lower target-to-total DNA ratios. In contrast, DNA extracts from swift fox scat exhibited low target DNA yields, with only 38% (19 of 50) of the samples showing comparative target DNA amplification of at least 0.1 ng. With this information, we were able to estimate the amount of target DNA entered into PCR amplifications, and identify samples having target DNA below a lower threshold of 0.2 ng and requiring modification to genotyping protocols such as multiple tube amplification. Our results here also show that this approach can easily be adapted to other species where faeces are the primary source of DNA template.