剪贴法设计条纹动感投影片的工艺

根据实践经验,为剪贴法设计条纹动感投影片建立了合理的工艺流程,介绍了它的一些基本方法．     

冷刺激下副溶血弧菌基因转录表达变化分析

目的:应用全基因组DNA芯片技术分析从37℃转入10℃后冷刺激作用下副溶血弧菌基因转录表达变化。方法:10℃低温分别作用于副溶血弧菌20、40和60min后,收集以上3个时间点和正常对照组的菌体,提取RNA,进行一个连续的时相分析。在每一个时间点的细菌基因表达分别与对照组(冷处理前37℃生长状态)相比较,获取每个时间点和正常对照组的基因转录表达上下调的倍数或者其对数值,以荧光信号的变化倍数值为2作为一个临界点,结合SAM分析软件选取基因变化倍数较大的数值。结果:在冷刺激后20、40、60min3个时间点可以检测到大量的转录水平发生明显变化的基因,对于这些发生变化的基因,根据各个时间点的变化规律,进行了分簇归类分析,共分为9簇,每一个簇都存在一种独特的时间依赖的表达模式。在每个相关的时间点,代谢相关的基因以下调为主。面对突然而急剧的温度变化,细胞膜相关的代谢基因发生了明显的重塑。结论:通过对体外低温环境下副溶血弧菌的比较转录谱分析,得到了在低温条件下基因转录发生明显变化的细胞代谢和毒力因子等相关基因,为副溶血弧菌基因转录调控网络的研究提供了一定的理论依据。     

Mitochondrial DNA evidence for the 19th century introduction of African honey bees into the United States

Since the introduction of an African subspecies into Brazil in the mid-1950's¹, descendent Africanized honey bees (Apis mellifera L.) have spread throughout the Neotropics and into temperate North America. Restriction enzyme analysis of 422 feral honey bee colonies collected from non-Africanized areas in the southern United States revealed that over 21% of them had mitochondrial DNA (mtDNA) derived from a European race established in North America by the 17th century, 77% of them had mtDNA common in honey bees maintained by beekeepers and about 1% exhibited African mtDNA. Further analysis revealed that the African mtDNA was derived from a north African subspecies imported to the US in the 19th century.     

Maternal vitamin A restriction alters biochemical development of the brain in rats

Summary The biochemical development of the fetal brain in relation to maternal vitamin A restriction was studied in rats. The vitamin A status of pregnant rats was varied by supplying low, medium and adequate amounts (6, 40, and 100 g retinol/day/kg body weight, respectively) of vitamin A during pregnancy and suckling. The maternal vitamin A restriction caused an altered brain development in terms of tissue weight, DNA, RNA and protein levels, and biosynthesis of DNA and protein from [³H]-thymidine and [³H]-leucine, respectively. A dose-dependent effect of maternal vitamin A restriction on the metabolism of DNA, RNA and protein was noticed in the developing fetal brain of rats.     

MGB probe assay for rapid detection of mtDNAl1778 mutation in the Chinese LHON patients by real-time PCR

Objective： Leber＇s hereditary optic neuropathy （LHON） is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA （mtDNA）. Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder （MGB） probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction （PCR）. Methods： Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA 1 1778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones. Results： All 48 LHON patients and their maternal relatives were positive for rntDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min. Conclusion： This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.     

Implications of ancient DNA for phylogenetic studies

The utility of DNA sequence characters from fossil specimens is examined from a phylogenetic perspective. Four ways that fossil characters can alter phylogenetic hypotheses are discussed. Two empirical examples and a third hypothetical example concerning amber-preserved insects are presented to illustrate these phenomena. Fossil DNA sequences as characters will be affected by the problem of missing data and missing taxa. In general, cladogram accuracy will be more greatly affected by missing taxa and cladogram resolution will be affected more acutely by missing data. Due to these points, an examination of the importance of the phylogenetic question being addressed, the utility of the fossil DNA sequences and the rarity of the fossil should be considered before damage of a fossil is undertaken.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

黑腹果蝇NDA修复功能缺陷型细胞系的建立及其培养特性

通过低温(18℃)培养方法,成功地建立了黑腹果蝇DNA修复功能缺陷型细胞系。该细胞系的特点是只能进行开放培养而不能进行封闭培养,为研究真核生物细胞DNA修复机理提供了一个有用的实验材料。     

DNA序列分类的统计分析

采用系统聚类分析方法，对巳知类别的20种DNA序列中4种碱基的含量及各碱基之间的相关性进行了统计分析，发现不同类别的DNA序列中减基的排列具有明显的规律性，由此建立了一种DNA序列分类的方法，并运用统计分析对这种分类方法的合理性给予了检验．     

Geographic populations of the medfly may be differentiated by mitochondrial DNA variation

Restriction enzyme cleavage sites of mitochondrial DNA (mtDNA) from the Mediterranean fruit fly were found to vary among introduced populations in the Neotropics. The survey included samples from 15 established natural populations and 5 laboratory cultures from Hawaii, Central America, South America and West Africa and samples from recent California infestations (1989, 1991). Based on restriction fragment length polymorphisms from 2 enzymes, Hawaii is an unlikely source for the 1989 and 1991 California infestations. Interpopulational variation in mtDNA demonstrates the potential for the technique to trace the process of colonization (geographic spread) by this insect.