应用RAPD技术对大熊猫分类地位的探讨

采用ＰＣＲ和Ｓｏｕｔｈｅｒｎ杂交等方法对大熊猫与小熊猫、马来熊、浣熊等共有的一条１．３ｋｂ的ＲＡＰＤ产物片段进行了初步分析。研究结果显示,来自于大熊猫的此共有片段可能为一重复序列,并且其内部含有多个随机引物ＡＢ１－０８的结合位点。以此来自于大熊猫的１．３ｋｂ片段为探针进行杂交,发现马来熊ＲＡＰＤ产物中的对应片段显示了很高的同源性,而小熊猫和浣熊ＲＡＰＤ产物则无相应的杂交带。这暗示,从分类地位上来看,大熊猫与熊科马来熊的亲缘关系应更近于与小熊猫和浣熊的亲缘关系,应与马来熊一样划分为熊科。本研究为大熊猫分类地位的确定提供了又一分子生物学证据     

熊超科的分子系统发生研究

有关熊超科的系统发生与演化存在很多争议。本工作测定了黑足鼬、南方海狮和非洲猎犬线粒体细胞色素ｂ、１２ＳｒＲＮＡ、ｔＲＮＡｔｈｒ和ｔＲＮＡＰｒｏ基因片段的ＤＮＡ序列。结合我们过去确定的熊超科其他动物的ＤＮＡ序列,我们采用简约法和距离法构建了较为系统的熊超科分子系统树。结果表明：浣熊科与鼬科、熊科与大熊猫科分别具有较近的共同祖先;鳍脚类不是一个独立的目,它们应归入熊超科;鳍脚类可能是单起源的,它们与浣熊科和鼬科的关系较与其他科更紧密;小熊猫科与其他科的关系并不密切。     

知识果味派

<正>小熊猫是小的大熊猫吗?当然不是!它们是完全不同的两种动物。从动物分类上来说,小熊猫和浣熊的亲缘关系比较近,大熊猫和黑熊的亲缘关系比较近。同样的,小独角犀不是小的大独角犀,它们也是两种不同的动物。小独角犀也叫爪哇犀,大独角犀又称印度犀。     

大熊猫,小熊猫,浣熊等五种动物毛的扫描电镜结构比较研究

本文对大熊猫、黑熊、棕熊、小熊猫和浣熊等动物毛的扫描电镜结构进行了比较研究,并对其主要鳞片类型的差异进行了ｔ检验。结果表明：大熊猫、黑熊、棕熊在毛的微观结构上与小熊猫、浣熊存在明显差异。     

蓝藻OscilatoriaceaerDNA16S-23S基因间隔区的序列分析及其分类学意义

采用末端终止法对蓝藻类颤藻科Ｏｓｃｉｌａｔｏｒｉａｓｐ．ｒＤＮＡ１６Ｓ－２３Ｓ基因间隔区进行了序列测定,获得了Ｏｓｃｉｌａｔｏｒｉａｓｐ．ｒＤＮＡ基因间隔区４２７个核苷酸,其中包含１个异亮氨酸ｔＲＮＡ基因（ｔＲＮＡＩｌｅ）。并通过计算机联网从国际分子生物学数据弹库中获取颤藻科其它种的ｒＤＮＡ基因间隔区序列,通过比较分析,从分子水平对颤藻科Ｏｓｃｉｌａｔｏｒｉａｃｅａｅ属间的某些分类学问题进行了讨论,并根据序列中核苷酸差异值探讨了颤藻科属间界定的分子标准。提出了ｒＤＮＡ基因间隔区是良好的分子标记,可用于“赤潮”或“水华”蓝藻专一性核酸分子探针的研制     

犬瘟热减毒疫苗对小熊猫安全性与免疫原性研究

犬瘟热病毒(canine distemper virus,CDV)是副粘病毒科麻疹病毒属成员,与麻疹病毒和牛瘟病毒亲缘关系很近,其基因组为不分节、非重叠的负链RNA,长15 960bp,由6个基因组成,编码N、P、M、F、H、L蛋白.此外,还包括由P基因编码的V和C蛋白[1].由CDV感染引起的犬瘟热,是一种急性、高度接触性传染病,主要引起犬科、鼬科和浣熊科动物的犬瘟热(CD),但是伴随生态环境的变化和犬瘟热病毒对动物流行病因素的适应,它的自然感染宿主范围在不断扩大,危害也越来越大.大熊猫(Ailuropoda melanoleuca)是我国特产的世界级珍稀濒危动物,由于大熊猫具有极高的科研和观赏价值,成了世界人民保护自然资源的象征.近年来有大熊猫发生CD的报道[2],研究大熊猫CD的预防是必要的.小熊猫作为浣熊科的动物,对CDV非常敏感[3].本研究通过犬瘟热减毒疫苗对小熊猫安全性与免疫原性研究,探讨小熊猫作为评价犬瘟热弱毒疫苗对大熊猫的安全性与免疫原性动物模型的可能性.     

我国汉族和彝族rRNA基因多态性的研究

人类ｒＤＮＡ顺序既偏偏保守性积蓄高的转录区,又有变异性较大的不转录间隔区。随着对ｒＤＮＡ结构的不断认识,发现对不转录间隔区特征的研究在分子群体遗传学上具有重要的价值。我们运用ｒＤＮＡ基因探针和Ｓｏｕｔｈｅｒｎ印迹法对甸汉族和彝族ｒＲＮＡ基因３＇端不转录间隔区作多态性分析,ＲＦＬＰ特征揭示两个民族有广泛的共性,又有各自的特点,这一遗传标志对少数民族群体遗传学乃至我国民族的迁移和演化的研究有重要意义。     

蓝藻Oscillatoriaceae rDNA16S—23S基因间隔区的序列分析及其分 …

采用末端终止法对蓝藻类颤藻科Ｏｓｃｉｌｌａｔｏｒｉａ ｓｐ．ｒＤＮＡ １６Ｓ－２３Ｓ基因间隔区进行了序列测定,获得了Ｏｓｃｉｌｌａｔｏｒｉａ ｓｐ．ｒＤＮＡ基因间隔区４２７个核苷酸,其中包含１个异亮氨酸ｔＲＮＡ基因。并通过计算机联网从国际分子生物学数据弹库中获取颤藻科其它种的ｒＤＮＡ基因间隔区序列,通过比较分析,从分子水平对颤藻科Ｏｓｃｉｌｌａｔｏｒｉａｃｅａｅ属间的某些分类学问题进行了讨论,并根     

二种淡水微囊藻rDNA16S-23S基因间隔区的序列测定与分析

本研究采用ＰＣＲ及序列测定的方法,对我国淡水铜绿微囊藻有毒株和另一低毒的种类惠氏微囊藻（Ｍ５７４）ｒＤＮＡ１６Ｓ－２３Ｓ基因间隔区进行了序列的测定和分析,结果表明：ｒＤＮＡ１６Ｓ－２３Ｓ基因间隔区可以作为一个精细且稳定的指标,用一微囊藻的分类和鉴定。并从分子水平提出了同囊藻与惠氏微囊藻在种系有较近缘的关系,本文首次对微落属ＭｉｃｒｏｃｙｓｔｉｓｒＤＮＡ基因间隔区全序列作了报道。为微囊藻属的鉴定及系     

应用兔抗大熊猫免疫球蛋白G血清探讨大熊猫的分类地位

从大熊猫血清中纯化出免疫球蛋白(IgG),以此作为抗原免疫家兔,获得兔抗大熊猫IgG血清。以黑熊、小熊猫、狗、猫等动物血清为抗原,兔抗大熊猫IgG 血清为抗体．进行了免疫扩散和微量免疫电泳实验。 实验结果表明,收集的食肉目动物：黑熊、小熊猫、狗、猫的血清都可与兔抗大熊猫GIg血清进行沉淀反应,其中尤以黑熊的反应最强且与大熊猫的反应沉淀线完全融合;小熊猫、狗、猫反应较弱且融入大熊猫反应沉淀线后形成树板状。从此看出大熊猫lgG 与黑熊的IgG最相似,从亲缘关系上讲,二者更为接近,大熊猫反应属熊科。     

MOLECULAR GENETIC-DISTANCE ESTIMATES AMONG THE URSIDAE AS INDICATED BY ONE- AND TWO-DIMENSIONAL PROTEIN ELECTROPHORESIS

Evolutionary relationships among eight species of Ursidae (including the giant panda) relative to two Procyonidae species (raccoon and red panda) were estimated based on the extent of electrophoretic variation of 289 radiolabelled fibroblast proteins resolved by two-dimensional gel electrophoresis and among 44 isozyme loci resolved by one-dimensional electrophoresis. Allelic differences among these species were converted to genetic distances, and phenetic trees were constructed. In addition, the electrophoretic data were coded as unit characters, and minimum-length trees were derived based on the Wagner method using maximum parsimony. Regardless of the tree-building method employed, the data sets agreed on the following branching sequence: between 22.4 and 32.3 million years (MY) ago, the ancestors of the procyonids and the ursids split into two lineages. Within 10 MY, the red panda split from the line that led to the raccoon. An ancestor of the giant panda split from the ursid line 18–22 MY ago, and the South American spectacled bear split from the line leading to ursine bears 10.5–15.0 MY B.P. A group of six closely related ursine bears (brown bear, polar bear, Asiatic black bear, Malayan sun bear, American black bear, and sloth bear) diverged from a common ancestor during the past 4–8 MY. Much of this ursine radiation was not resolved by our results, with the exception of a recent (2–3 MY B.P.) divergence of brown bear and polar bear. The topological concordance of the data sets from one- and two-dimensional electrophoresis supports the usefulness of these procedures for evolutionary inference and provides additional precision to the reconstruction of divergence nodes of this carnivore group.     

大熊猫等五种食肉动物血清蛋白和LDH同工酶盘电泳比较

大熊猫(Ailuropoda melanoleuca)是世界珍稀动物之一,近年来,兽类科学工作者对它进行了多方面的研究,也包括生化方面的工作(潘文石等,1982;Sarich,1973),但与其他食肉哺乳动物相比研究较少,而此又为探讨其分类地位所需。本文应用聚丙烯酰胺凝胶盘电泳,对大熊猫、小熊猫(AilurSs fulgens)、黑熊(Selenarctos thibetanus)、家猫及犬等5种食肉动物的血清蛋白和LDH同工酶进行比较分析,目的在于了解这5种动物的血清蛋白图象和LDH同工酶酶谱以及它们之间的谱型、相对活力、迁移率的异同,进而讨论大熊猫的分类地位。     

大熊猫、小熊猫和黑熊血红蛋白的组份分离、结晶及氨基酸组成分析

 聚丙烯酰胺凝胶电泳和羧甲基纤维素柱层析都表明大熊猫(Ailuropodmelanoleuca)、小熊猫(Ailurus fulgens)和黑熊(Selenarctos thibetanus)的血红蛋白各有两个组份。在近似的条件下得到了它们的晶体。三种动物血红蛋白的氨基酸组成相似,而且N-末端氨基酸都是Val。有待进一步测定全部氨基酸顺序,以揭示它们之间的异同和亲缘关系。     

The brain of the lesser panda Ailurus fulgens: A quantitative approach

A quantitative analysis of the brain of the lesser panda Ailurus fulgens was carried out, using both ratios and progression indices. In a general way, the lesser panda appears to be an intermediate form between the giant panda and the raccoon. Relations between brain size or brain component size, and life-habits, suggest the same conclusion. The traditionally assumed closeness of both pandas implied in the vernacular names, is questioned. A comprehensive neuroethological work on the comparative organization of the brain in all ursoid species is desirable, as a good case in fundamental functional morphology.     

Chromosomal rearrangements and karyotype evolution in carnivores revealed by chromosome painting

Chromosomal evolution in carnivores has been revisited extensively using cross-species chromosome painting. Painting probes derived from flow-sorted chromosomes of the domestic dog, which has one of the most rearranged karyotypes in mammals and the highest dipoid number (2n=78) in carnivores, are a powerful tool in detecting both evolutionary intra- and inter-chromosomal rearrangements. However, only a few comparative maps have been established between dog and other non-Canidae species. Here, we extended cross-species painting with dog probes to seven more species representing six carnivore families: Eurasian lynx (Lynx lynx), the stone marten (Martes foina), the small Indian civet (Viverricula indica), the Asian palm civet (Paradoxurus hermaphrodites), Javan mongoose (Hepestes javanicas), the raccoon (Procyon lotor) and the giant panda (Ailuropoda melanoleuca). The numbers and positions of intra-chromosomal rearrangements were found to differ among these carnivore species. A comparative map between human and stone marten, and a map among the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis), stone marten and human were also established to facilitate outgroup comparison and to integrate comparative maps between stone marten and other carnivores with such maps between human and other species. These comparative maps give further insight into genome evolution and karyotype phylogenetic relationships among carnivores, and will facilitate the transfer of gene mapping data from human, domestic dog and cat to other species.     

凉山山系大熊猫和黑熊适宜生境预测及重叠分析

研究同域物种的分布格局及重叠状况对物种的区域整合保护管理及区域生物多样性保护具有重要实践价值。本研究基于全国第四次大熊猫调查及长期野外调查数据, 利用MaxEnt模型预测了凉山山系两种同域分布的熊科动物——大熊猫(Ailuropoda melanoleuca)和黑熊(Ursus thibetanus)的适宜生境, 基于适宜生境预测结果, 分析了两个物种的生境需求因子、生境破碎化现状及重叠状况。结果显示: (1)大熊猫和黑熊的适宜生境分布格局相似, 主要分布在凉山山系的山脊地带, 适宜生境面积分别为1,383.84 km²和2,411.49 km²; (2)两个物种的适宜生境都较为破碎, 且存在一些隔离分布区, 相较而言, 黑熊适宜生境的连通性要优于大熊猫; (3)两个物种生态位重叠度较高(D = 0.654, I = 0.901), 适宜生境重叠面积为958.29 km², 分别占大熊猫和黑熊适宜生境总面积的69.25%和39.74%; (4)两个物种对环境因子的选择和响应表现出了相似性和差异性。相似性在于对两个物种生境分布影响最大的两个因子均为距居民点距离和海拔; 差异性在于对大熊猫生境分布影响次之的因子是植被类型和最冷季均温, 而黑熊的是年最大EVI指数和距道路距离。为了更有效地保护两个物种, 应加强对人类干扰的控制和植被的恢复, 对栖息地实行连通管理, 并建立多物种保护规划。     

Mandible of the giant panda (Ailuropoda melanoleuca) compared with other Chinese carnivores: functional adaptation

The aim of this study was to understand the mandible of the giant panda in morphometric terms to explore differences between the giant panda and other carnivores distributed in China, in terms of functional adaptation. Twelve mandibular variables were studied using bivariate (allometry) and multivariate (principal components analysis, PCA, and discriminant functional analysis, DFA) tools. When deviations were produced from allometric baselines consisting of all the species studied, the giant panda displayed a much more developed mandibular structure than the bear, leopard, and tiger. This may be related to its specific dietary preference for bamboo, which has very strong fibers. Results also indicate that the mandibular structure among carnivores mainly reflects the differences in their dietary preferences and functional adaptation. Three groups were found referring to dispersal profiles expressed by the first two axes of PCA and DFA: (1) the two panda species – the herbivorous carnivores; (2) the black bear – the omnivorous carnivore; and (3) the tiger and leopard – the hypercarnivores. Nevertheless, a significant separation between the two panda species was also found with the profiles displayed by the first and third axes of DFA. In addition to no close evolutionary relationship and phylogenetic development, a noticeable separation between the two panda species found in DFA analysis may be associated with their variation in consuming different parts of the bamboo plant: the giant panda feeds on stems and the red panda feeds on leaves.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 449–456.     

大熊猫和亚洲黑熊TNNC1 基因的克隆、表达与序列分析

慢肌肌钙蛋白C (Troponin C type 1,TNNC1)具有高度保守性,调控骨骼肌慢肌和心肌的收缩,影响肌蛋白的生成,从而可能导致动物肌肉的生长、进化和功能的差异。本研究以大熊猫和亚洲黑熊骨骼肌为材料,提取总RNA 和基因组DNA,运用RT-PCR 和Touch-down PCR 分别扩增出TNNC1 基因的cDNA 序列和结构基因序列,并且构建了含有TNNC1 cDNA 的重组表达载体,转化进入E. coli BL21 进行超表达研究。结果表明大熊猫TNNC1 基因的cDNA 片段长602 bp,包含一个编码161 个氨基酸的开放阅读框,其结构基因全长2 831 bp,包含6 个外显子和5 个内含子。亚洲黑熊TNNC1 基因的cDNA 片段长486 bp,亦包含一个编码161 个氨基酸的开放阅读框,其结构基因全长2 758 bp,同样包含6 个外显子和5 个内含子。该两个物种的TNNC1 基因与已报道的13种动物的TNNC1 基因具有很高的相似性。拓扑预测表明,大熊猫和亚洲黑熊TNNC1 蛋白有1 个蛋白激酶C 磷酸化位点,5 个酪蛋白激酶Ⅱ磷酸化位点,1 个N-豆蔻酰化位点,3 个EF 手性钙结合域及1 个N - 糖基化位点。将TNNC1 基因在大肠杆菌中表达发现TNNC1 蛋白与氮端多聚组氨酸标签蛋白(His6) 融合成大小为23. 5kD 左右的多肽,这与预期结果相一致。本研究结果为进一步深入探讨大熊猫和亚洲黑熊TNNC1 基因及蛋白的结构、功能和进化关系提供资料。