Foot-and-mouth disease virus VP1 protein fused with cholera toxin B subunit expressed in Chlamydomonas reinhardtii chloroplast

A Chlamydomonas reinhardtii chloroplast expression vector, pACTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed and transfered to the chloroplast genome of C. reinhardtii by the biolistic method. The transformants were identified by PCR, Southern blot, Western blot and ELISA assays after selection on resistant medium and incubation in the dark. The CTBVP1 fusion protein was expressed in C. reinhardtii chloroplast and accounted for up to 3% of the total soluble protein. The fusion protein also retained both GM1-ganglioside binding affinity and antigenicity of the FMDV VP1 and CTB proteins. These experimental results support the possibility of using transgenic chloroplasts of green alga as a mucosal vaccine source.     

用指纹图谱评价环境样本DNA的提取效果

研究了DNA指纹图谱技术在评价环境样本DNA提取效果方面的应用。采用 3种DNA提取方法提取垃圾渗滤水和活性污泥中的微生物DNA ,并用ARDRA和RISA图谱加以评价。实验结果表明 ,RISA图谱分析法可以作为评估DNA提取效果的有效手段。     

同位素示踪技术在丛枝菌根真菌生态学研究中的应用

丛枝菌根(arbuscular mycorrhizal,AM)真菌是生态系统中重要的土壤微生物之一。AM真菌菌丝体网络是由AM真菌菌丝体在土壤生态系统中连接两株或两株以上植物根系所形成的菌丝体网络。随着菌根学研究的深入,如何直观的揭示AM真菌的生态学功能已经成为相关领域关注的热点问题。研究发现,利用同位素示踪技术可以开展AM真菌与宿主植物对土壤矿质营养的吸收、转运等方面的研究,以及菌丝体网络对不同宿主植物之间营养物质的分配研究和AM真菌在生态系统生态学中的功能研究。基于此,为了阐明同位素示踪技术在AM真菌研究中的价值,围绕菌根学最新研究进展,系统回顾了利用同位素示踪技术探究AM共生体对不同元素吸收和转运的机制、同位素示踪技术在AM真菌菌丝体网络研究中的价值和利用同位素示踪技术研究AM真菌在生态系统中的功能,为AM真菌生态学功能的研究提供理论基础,并对本领域未来的研究方向和应用前景进行展望。     

干旱和湿润生境下辽东栎群体遗传结构及其适应意义的初步研究

应用同工酶分析方法,测定北京市东灵山区两个分别代表干旱和湿润生境的辽东栋（ＱｕｅｒｃｕｓｌｉａｏｔｕｎｇｅｎｓｉｓＫｏｉｚ．）群体的遗传结构。共分析统计了１３ 个酶系统３０ 个位点。结果表明：辽东栎群体内部存在丰富的遗传变异（多态位点百分率为８６．６％ ,等位基因平均数为２．２５）。两群体遗传结构的相似性程度很高（Ｄ＝ ０．０２９, ＧＳＴ＝ ０．０４８）;但在个别位点上仍存在较大差异,这些差异的产生可能与对小生境的适应有关。对不同年龄段的初步分析结果显示,基因频率的动态变化可能有其适应意义     

Assessing Mitochondrial DNA Variation and Copy Number in Lymphocytes of ~2,000 Sardinians Using Tailored Sequencing Analysis Tools

DNA sequencing identifies common and rare genetic variants for association studies, but studies typically focus on variants in nuclear DNA and ignore the mitochondrial genome. In fact, analyzing variants in mitochondrial DNA (mtDNA) sequences presents special problems, which we resolve here with a general solution for the analysis of mtDNA in next-generation sequencing studies. The new program package comprises 1) an algorithm designed to identify mtDNA variants (i.e., homoplasmies and heteroplasmies), incorporating sequencing error rates at each base in a likelihood calculation and allowing allele fractions at a variant site to differ across individuals; and 2) an estimation of mtDNA copy number in a cell directly from whole-genome sequencing data. We also apply the methods to DNA sequence from lymphocytes of ~2,000 SardiNIA Project participants. As expected, mothers and offspring share all homoplasmies but a lesser proportion of heteroplasmies. Both homoplasmies and heteroplasmies show 5-fold higher transition/transversion ratios than variants in nuclear DNA. Also, heteroplasmy increases with age, though on average only ~1 heteroplasmy reaches the 4% level between ages 20 and 90. In addition, we find that mtDNA copy number averages ~110 copies/lymphocyte and is ~54% heritable, implying substantial genetic regulation of the level of mtDNA. Copy numbers also decrease modestly but significantly with age, and females on average have significantly more copies than males. The mtDNA copy numbers are significantly associated with waist circumference (p-value = 0.0031) and waist-hip ratio (p-value = 2.4×10^-5), but not with body mass index, indicating an association with central fat distribution. To our knowledge, this is the largest population analysis to date of mtDNA dynamics, revealing the age-imposed increase in heteroplasmy, the relatively high heritability of copy number, and the association of copy number with metabolic traits.     

黄河下游平原不同非农生境中植物多样性

自然、半自然等非农生境是区域农业景观的重要组成部分,对区域生物多样性保护具有重要意义。黄河下游平原区是典型的农业景观,农田是最主要的景观类型,林地、树篱等景观要素散布其中,为研究区内非农生境中生物多样性及其生态效应,采用典型样地法对区内典型农业景观中林地、树篱、田间道路和沟渠等主要非农生境中的植物群落进行调查研究。结果显示:区内植物组成以菊科、禾本科等为主;区内植物科的地理成分以世界分布和泛热带分布为主,各生境间有一定的差异,属的地理成分复杂,具有中国15个种子植物属分布型中的13个,以温带分布、世界分布和泛热带分布为主,总体上区内的植物组成以广布种为主,优势科属明显,特有种缺乏;各非农生境中的植物多样性存在一定的差异,林地和树篱生境具有较高的物种丰富度和多样性,显著高于田间道路,沟渠、林地和树篱生境中的物种均匀度和群落盖度均显著高于田间道路;β多样性分析表明田间道路生境中的群落组成分化程度在各样点间最大(β多样性指数最高);树篱、林地和田间道路等生境间群落相似性均较高,但其群落结构和优势种组成方面却存在显著的差异,沟渠作为一种特殊生境与其它生境间的群落相似性相对较低。研究表明,在黄河下游平原典型农业景观中,作为非农生境存在的林地和树篱在物种多样性维持中具有重要地位,沟渠为水生和湿生植物提供了庇护所,意义重大;各生境间高的群落相似性仅是物种组成名录相似性的反映,其空间格局和优势种群间差异明显,各生境植物群落的生态功能差异巨大。未来区内生物多样性的保护应重在生态系统过程、功能的加强以及生态系统服务的维持和提高,且需进一步在景观水平上探讨各非农景观要素的空间构型对其生态效益的影响机制及其调控和管理策略。     

西北产6种药用柴胡营养器官的比较解剖学研究

柴胡是伞形科柴胡属植物,据报道,全世界有150种左右,主要分布在北半球及亚热带地区,初步统计我国有30余种。据我们在西北五省调查,药用种类有21种之多,药用主流种类有柴胡Bupleurum chinesis DC.、狭叶柴胡B. scorzonerifolium Willd.、银州柴胡B. yinchowense Shan et Y. Li、小叶黑柴胡B. smithii Wolff. var. parvi-     

高效的植物DNA提取方法

利用液氮研磨植物幼芽,以苯酚─氯仿─异戊醇─核糖核酸酶法提取了大豆、菜豆、玉米、高梁等几种农作物的总ＤＮＡ。所提取的ＤＮＡ经岛津ＵＶ－２６５紫外分光光度计检测及０．６％的琼脂糖凝胶电泳,结果证明：该方法所提取的植物总ＤＮＡ纯度高,片段长度整齐,约５０ｋｂ左右,符合外源ＤＮＡ导入作物的要求,并且ＤＮＡ收率很高。