EoNPV的限制性酶切图谱及polyhedrin基因和egt基因的定位

用８种限制性内切酶酶解茶尺蠖Ｅｃｔｒｏｐｉｓｏｂｌｉｑｕａ核型多角体病毒（ＥｏＮＰＶ）基因组ＤＮＡ，获得大于０．８８ｋｂ的片段数分别为７、３８、１９、１７、１６、９、３２和１４．由此统计，基因组平均大小大于１１８．５ｋｂ，即Ｍｒ约大于７８．６×１０６．用ＢｍＮＰＶ的多角体蛋白（ｐｈ）基因和ＡｃＭＮＰＶ的ｅｇｔ基因为探针，应用Ｓｏｕｔｈｅｒｎ杂交方法，将ＥｏＮＰＶ的ｐｈ基因定位于ＢａｍＨＩＡ、ＣｌａＩＤ、ＥｃｏＲＩＭ、ＥｃｏＲＶＬ、ＨｉｎｄⅢＦ、ＰｓｔＩＡ、ＳａｌＩＨ和ＸｈｏＩＢ片段上，将ｅｇｔ基因定位于ＢａｍＨＩＡ、ＣｌａＩｌ、ＥｃｏＲＩＫ、ＥｃｏＲＶＡ、ＨｉｎｄⅢＨ、ＰｓｔＩＡ、ＳａｌＩＢ、ＸｈｏＩＩ片段上．通过克隆和酶切鉴定ＥｃｏＲＩＭ和ＥｃｏＲＶＬ片段，测定ｐｈ基因部分序列，并以ＥｃｏＲＩＭ为探针对基因组酶切印迹进行杂交，制作了ＥｏＮＰＶｐｈ基因和ｅｇｔ基因区的酶切图谱，推定ｅｇｔ基因保守区位于ｐｈ基因上游约４．５５ｋｂ处．     

邻苯二酚1,2-双加氧酶(C12O)基因的定位、克隆和表达

射性同位素标记ｔｆｄＣ基因片段作探针,Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交定位Ｌ１菌株的邻苯二酚１,２双加氧酶基因位于ＰｓｔⅠ的Ｉ片段和ＢａｍＨⅠ的Ｍ和Ｎ片段．低熔点琼脂糖回收ＰｓｔⅠ的Ｉ段,直接克隆至表达载体ｐＫＴ２３０上,获得重组子ｐＫＴ２３０ｐ．重组子转化不具开环酶活性的甲胺磷降解菌Ｐ２,获得高效的Ｃ１２Ｏ酶活     

BACILLUS SP. BT-7木聚糖酶基因克隆与鉴定

报道了从一株有较高半纤维素酶活性的Ｂａｃｉｌｕｓｓｐ．ＢＴ７克隆内切木聚糖酶基因的结果．以大肠杆菌ＸＬ１ｂｌｕｅ为宿主菌,采用水解圈检测法,在含有ＲＢＢ木聚糖（４氧甲基Ｄ葡萄糖苷Ｄ木聚糖Ｒｅｍａｚｏｌ亮蓝Ｒ）的平板上分离到能水解ＲＢＢ木聚糖的阳性克隆３个,对它们进行的限制酶作图和Ｓｏｕｔｈｅｒｎ杂交分析表明,它们属于３种不同的内切木聚糖酶基因     

用简并引物扩增与克隆D.radiodurans超氧化物歧化酶和过氧化氢酶基因片段

依据超氧化物歧化酶（ＳＯＤ） 和过氧化氢酶（Ｃａｔ） 的保守性氨基酸序列,设计简并引物,自Ｄ．ｒａｄｉｏｄｕｒａｎｓ 基因组ＤＮＡ扩增并克隆了ＳＯＤ和Ｃａｔ 基因片段．ＳＯＤ 和Ｃａｔ 基因片段分别长４５３ ｂｐ 和６７３ ｂｐ,各编码１５１和２２４ 个氨基酸,氨基酸序列与不同生物来源的ＳＯＤ或Ｃａｔ 具有高度同源性     

人G—CSF转基因鼠系与正常鼠系比较研究

利用人ＧＣＳＦ基因组基因和ｃＤＮＡ基因作为目的片段，将其受控于２．６ｋｂ的小鼠乳清酸蛋白（ＷＡＰ）基因的调控区下，通过显微注射法获得了两只整合有人ＧＣＳＦ转基因小鼠，通过繁殖建立了稳定的转基因系．一些表型参数测定表明，转基因鼠与正常鼠无明显差别，为对转基因动物进行安全性评价提供了依据     

大熊猫肌肉氨基酸含量的初步研究

测定了３只大熊猫的头部、背部和腿部等１０个肌肉样品中１７种氨基酸（ＡＡ）含量,并分析了各样品ＡＡ与Ｌｙｓ含量的比值．结果表明：对同一只大熊猫,不同部位的肌肉样品中各ＡＡ含量无明显差异;对同一个肌肉样品,１７种ＡＡ含量比较,Ｇｌｕ含量最高,Ａｓｐ、Ａｒｇ、Ｌｙｓ、Ｌｅｕ、Ａｌａ、Ｇｌｙ、Ｖａｌ、Ｐｒｏ、Ｔｈｒ、Ｐｈｅ等含量较高;不同年龄大熊猫肌肉ＡＡ含量有差异,除极少数ＡＡ外,未成熟雄性大熊猫肌肉ＡＡ含量高于成熟和老龄雌性大熊猫,且有显著性差异（Ｐ＜０．０５或Ｐ＜０．０１）;成熟雌性大熊猫肌肉中,除Ｐｒｏ外,其余所有ＡＡ含量与其年龄无显著差异（Ｐ＞０．０５）;除极少数ＡＡ外,不同年龄、性别大熊猫肌肉样品中各ＡＡ与Ｌｙｓ含量的比值无明显差异     

黑曲霉(ASPERGILLUS NIGER)SG株三磷酸甘油醛脱氢酶基因的分离与初步鉴定

黑曲霉（Ａｓｐｅｒｇｉｌｕｓｎｉｇｅｒ）在工业生产和科学研究中占有重要地位，因为许多黑曲霉菌株能产生有机酸和酶制剂，并能分泌大量蛋白质到细胞外［１，６］．三磷酸甘油醛脱氢酶（Ｇｌｙｃｅｒａｌｄｅｈｙｄｅ３ｐｈｏｓｐｈａｔｅｄｅｈｙｄｒｏｇｅｎａ...     

粗壮女贞中的新单萜配糖体(英)

从粗壮女贞（Ｌｉｇｕｓｔｒｕｍ ｒｏｂｕｓｔｕｍ）叶子中分离鉴定了４ 个新的单萜配糖体, 命名为粗壮女贞甙Ｄ、Ｇ、Ｈ和Ｌ． 通过波谱及化学方法,它们分别鉴定为香叶醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（１） ,６羟基３,７二甲基２Ｅ,７葵二烯醇３′ＯαＬ吡喃鼠李糖基４′反式香豆酰基βＤ吡喃葡萄糖甙（２） ,６羟基３,７二甲基２Ｅ,７葵二烯醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（３） 以及６,７二羟基３,７二甲基２Ｅ葵烯醇３′ＯαＬ吡喃鼠李糖基４′顺式香豆酰基βＤ吡喃葡萄糖甙（４） ． 此外,还从该植物得到了７ 个已知化合物,芹菜素（５）、ｃｏｓｍｏｓｉｉｎ（６） 、ｒｈｏｉｆｏｌｉｎ（７） 、山梨糖醇（８）、阿克苷（９） 、β谷甾醇（１０） 和齐墩果酸（１１） ．     

用简并引物扩增与克隆D.radiodurans超氧化物歧化酶和过氧化 …

依据超氧化物歧化酶（ＳＯＤ）和过氧化氢酶（Ｃａｌ）的保守性氨基酸序列,设计简并引物,自Ｄ．ｒａｄｉｏｄｕｒａｎｓ基因组ＤＮＡ扩增并克隆了ＳＯＤ和Ｃａｔ基因片 Ｃａｔ基因片段分别长４５３ｂｐ和６７３ｂｐ,各编码１５１和２２４个氨基酸,氨基酸序列与不同生物来源的ＳＯＤ或Ｃａｔ具有高度同源性。     

固定CO2基因工程菌的构建

构建了含有ＲｕｂｉｓＣＯＦｏｒｍ ＩＩ基因的可转化大肠杆菌紫色非硫杆菌穿梭质粒（ＰＭＰＢ２）,将其转化紫色非硫杆菌野生型Ｒｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｐａｌｕｓｔｒｉｓ Ｎｏ．７ 和紫色非硫杆菌ＲｕｂｉｓＣＯＦｏｒｍ Ｉ缺陷型ＲｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｐａｌｕｓｔｒｉｓＮｏ．７ＤＦＩ,获得２ 株ＲｕｂｉｓＣＯ基因工程菌ＭＧ１１ 和ＭＧ１４,二者的ＲｕｂｉｓＣＯ 酶活性比对照菌分别提高７６ ．７％ 和８３．３％ ．连续传４０ 代表明,重组质粒在受体菌中是较稳定的     

圈养大熊猫乳汁营养成分分析及育幼营养对策初探

连续测定了１只１０ａ大熊猫产后１ｍｏ内１２个乳汁样品中水份、粗蛋白（ＣＰ）、粗脂肪（ＥＥ）、粗灰分（ＣＡ）、乳糖（Ｌａｃ）、１７种氨基酸（ＡＡ）、１０种矿物元素、６种维生素的含量．结果表明，大熊猫产后前６ｄ乳汁中多数营养素含量与６ｄ后含量差异明显．产后ｄ１乳汁中ｗ（ＣＰ）＝６．３４％，ｄ５～１５在５％左右；ｗ（ＥＥ）在２．５０％～７．３３％范围内变动；乳糖含量皆在１％左右；大多数维生素和微量元素含量远高于常见的家畜乳和人乳中相应成分的含量．在分析大熊猫乳汁营养成分含量特点的基础上，探讨了育纳营养对策．     

Application of least-cost path model to identify a giant panda dispersal corridor network after the Wenchuan earthquake—Case study of Wolong Nature Reserve in China

With growing levels of human-activity and frequent natural disturbances throughout the world, it is increasingly important that both research and management efforts take into account the widespread landscape fragmentation and its consequences for biodiversity conservation. The magnitude 5.12 Wenchuan earthquake in China caused dramatic impacts on giant panda (Ailuropoda melanoleuca) habitat in the nature reserves within Minshan and Qionglai mountains. With the combined stresses of the natural disaster and the extensive human activities during postquake reconstruction, giant panda habitat in this region may become more fragmented in the future. In order to preserve the giant panda population after the earthquake and protect the species against habitat fragmentation, this article explores a method of identifying giant panda migration corridors involving habitat suitability assessments and a least-cost path model. Focusing on postquake Wolong Nature Reserve, our results demonstrate that it contains 430.3 km² of suitable habitat (21.1% of total area), 463.8 km² of marginally suitable habitat (22.8%) and 1141.9 km² of unsuitable habitat (50.1%). We further show that several giant panda dispersal corridors exist in the reserve, including four corridor groups that cross the provincial highway and five corridors that do not intersect areas of human activity. This study will contribute to management and conservation efforts in Wolong Nature Reserve and beyond after the Wenchuan earthquake.     

Proposed Conservation Landscape for Giant Pandas in the Minshan Mountains,China

Abstract: The giant panda (Ailuropoda melanoleuca), is one of the world's most endangered species. Habitat loss and fragmentation have reduced its numbers, shrunk its distribution, and separated the population into isolated subpopulations. Such isolated, small populations are in danger of extinction due to random demographic factors and inbreeding. We used least‐cost modeling as a systematic approach to incorporate satellite imagery and data on ecological and behavioral parameters of the giant panda collected during more than 10 years of field research to design a conservation landscape for giant pandas in the Minshan Mountains. We identified 8 core habitats and 4 potential linkages that would link core habitats CH3, CH4, and CH5 with core habitats CH6, CH7, and CH8. Establishing and integrating the identified habitats with existing reserves would create an efficient reserve network for giant panda conservation. The core habitats had an average density of 4.9 pandas/100 km² and contained approximately 76.6% of the giant panda population. About 45% of the core habitat (3245.4 km²) existed outside the current nature reserves network. Total estimated core habitat decreased between 30.4 and 44.5% with the addition of residential areas and road networks factored into the model. A conservation area for giant panda in the Minshan Mountains should aim to ensure habitat retention and connectivity, improve dispersal potential of corridors, and maintain the evolutionary potential of giant pandas in the face of future environmental changes.     

Characteristics that make trophy hunting of giant pandas inconceivable

In November 1928, Theodore Jr. and Kermit Roosevelt led an expedition to China with the expressed purpose of being the first Westerners to kill the giant panda (Ailuropoda melanoleuca). The expedition lasted 8 months and resulted in the brothers shooting a giant panda in the mountains of Sichuan Province. Given the concurrent attention in the popular press describing this celebrated expedition, the giant panda was poised to be trophy hunted much like other large mammals around the world. Today, however, the killing of giant pandas, even for the generation of conservation revenue, is unthinkable for reasons related to the species itself and the context, in time and space, in which the species was popularized in the West. We found that the giant panda's status as a conservation symbol, exceptional charisma and gentle disposition, rarity, value as a nonconsumptive ecotourism attraction, and endemism are integral to the explanation of why the species is not trophy hunted. We compared these intrinsic and extrinsic characteristics with 20 of the most common trophy-hunted mammals to determine whether the principles applying to giant pandas are generalizable to other species. Although certain characteristics of the 20 trophy-hunted mammals aligned with the giant panda, many did not. Charisma, economic value, and endemism, in particular, were comparatively unique to the giant panda. Our analysis suggests that, at present, exceptional characteristics may be necessary for certain mammals to be excepted from trophy hunting. However, because discourse relating to the role of trophy hunting in supporting conservation outcomes is dynamic in both science and society, we suspect these valuations will also change in future.     

大熊猫幼仔口腔分泌物中检出聚团肠杆菌和微球菌

大熊猫在圈养条件下 ,配种难、受孕难、幼仔成活更难已众所周知[1,2 ] .本文报告了一只患病大熊猫幼仔的静脉血、尿液和粪便的常规检测和细菌学分离、鉴定 ,并从该幼仔口腔分泌物中检出聚团肠杆菌和微球菌的结果 .以为大熊猫幼仔疾病的检测、诊断和治疗积累基础资料 .1　实验室检查大熊猫幼仔一只 ,1998年 9月出生于成都大熊猫繁育研究基地 ,初生重 110 ｇ左右 ,出生第 3天 ,出现体温间隙性增高 ,拉血便 ,食欲减少 ,烦躁不安 ,精神状态较差 .1.1　常规检查用卡介苗注射用的针头采集颈静脉血作血常规检验 ,其结果 :白细胞总数 :2 3.6× 10 9/…     

Patterns of Genetic Diversity in Remaining Giant Panda Populations

Abstract: The giant panda ( Ailuropoda melanoleuca ) is among the more familiar symbols of species conservation. The protection of giant panda populations has been aided recently by the establishment of more and better-managed reserves in existing panda habitat located in six mountain ranges in western China. These remaining populations are becoming increasingly isolated from one another, however, leading to the concern that historic patterns of gene flow will be disrupted and that reduced population sizes will lead to diminished genetic variability. We analyzed four categories of molecular genetic markers (mtDNA restriction-fragment-length polymorphisms [RFLP], mtDNA control region sequences, nuclear multilocus DNA fingerprints, and microsatellite size variation) in giant pandas from three mountain populations (Qionglai, Minshan, and Qinling) to assess current levels of genetic diversity and to detect evidence of historic population subdivisions. The three populations had moderate levels of genetic diversity compared with similarly studied carnivores for all four gene measures, with a slight but consistent reduction in variability apparent in the smaller Qinling population. That population also showed significant differentiation consistent with its isolation since historic times. From a strictly genetic perspective, the giant panda species and the three populations look promising insofar as they have retained a large amount of genetic diversity in each population, although evidence of recent population reduction—likely from habitat loss—is apparent. Ecological management to increase habitat, population expansion, and gene flow would seem an effective strategy to stabilize the decline of this endangered species.     

卧龙大熊猫人工育幼技术的研究进展

简要回顾了大熊猫人工育幼的历史，总结了卧龙中国保护大熊猫研究中心在大熊猫人工育幼研究领域的发展。1991年全人工育幼使1只大熊猫弃仔存活160d，但之后连续几年人工育幼失败；1998年成功地将一只弃仔人工哺育2mo后，给不会带仔的母兽培训其带仔。1999年将2个双胞胎及1个三胞胎中的2仔通过人工育幼与母兽交替哺育存活，同时将2只自野外抢救回来的大熊猫幼仔人工哺育存活。创造了1a人工哺育8只幼仔存活的世界记录，大大提高了大熊猫幼仔的存活率。大熊猫人工育幼技术的关键在于；适宜的育幼环境，幼仔免疫力的获得，人工乳的配制，幼兽的疾病防治及正确的饲喂方式。现将卧龙大熊猫庆大熊猫人工育幼方面所取得的突破性进展总结如下。图2表2参6     

A Framework for Evaluating the Effects of Human Factors on Wildlife Habitat: the Case of Giant Pandas

Abstract: To address the complex interactions between humans and wildlife habitat, we developed a conceptual framework that links human factors with forested landscapes and wildlife habitat. All the components in the framework are integrated into systems models that analyze the effects of human factors and project how wildlife habitat would change under different policy scenarios. As a case study, we applied this framework to the Wolong Nature Reserve in Sichuan Province (southwestern China), the largest home of the giant panda ( Ailuropoda melanoleuca ). We collected ecological and socioeconomic data with a combination of various methods ( field observations, aerial photographs, government documents and statistics, interviews, and household surveys) and employed geographic information systems and systems modeling to analyze and integrate the data sources. Human population size has increased by 66% and the number of households in the reserve has increased by 115% since 1975, when the reserve was established. During the same period, the quality and quantity of the giant panda habitat dramatically decreased because of increasing human activities such as fuelwood collection. Systems modeling predicted that under the status quo, human population in the reserve would continue to grow and cause more destruction of the remaining panda habitat, whereas reducing human birth rates and increasing human emigration rates would lower human population size and alleviate human impacts on the panda habitat. Furthermore, our simulations and surveys suggested that policies encouraging the emigration of young people would be more effective and feasible than relocating older people in reducing human population size and conserving giant panda habitat in the reserve.     

黑斑蛙Dmrt1基因的克隆及在不同组织中的表达

Dmrt基因家族是新近发现的一个与性别决定相关的基因家族,该家族成员都含有一个新的具有DNA结合能力的保守基序——DM结构域.本文采用简并PCR技术,扩增并克隆了黑斑蛙基因组中的DM结构域,经序列分析,获得了Dmrt家族成员rnDmrt1;进一步根据获得的rnDmrt1序列,设计一对特异引物,采用RT-PCR技术对成蛙不同组织Dmrt1基因的表达进行了研究.结果发现,rnDmrt1只在精巢中特异表达,在卵巢、脑、肝、心、肾及肌肉组织中无表达,显示Dmrt1基因在性别决定和分化中有重要功能. 图4参9     

小麦高分子量谷蛋白亚基基因的PCR及AS-PCR标记

选用高分子量麦谷蛋白亚基(HMW-GS)在Glu-D1位点已知的 26个普通小麦品种,包括 15个 (TriticumaestivumL. )川育系列品种及其 11个亲本,根据其亚基Dx5和Dy10基因的特异序列设计引物.通过对Dx5基因的PCR扩增及对Dy10基因的AS-PCR扩增,结果表明,所设计的引物对Dx5和Dy10都能扩增出特异条带,而携有Dx5基因的材料同时也携有Dy10基因.说明用所设计的引物可快速有效的鉴定出普通小麦中是否带有优质的 5 10亚基.图 4参 15