Biotinylation of double stranded DNA after transamination

The bisulfite catalyzed transamination of cytidine and cytosine has been reported to be single strand specific, but local thermal instabilities of the DNA double helix, coupled with the extreme sensitivity of the Biotin-Avidin revelation methods, allows the random labelling of cytosines in d.s. DNA to detectable levels for those purposes where the overall label can be very low. We have evaluated the use of this reaction to prepare double stranded DNA molecules containing N4-aminoethyl-cytosine (4-aeC). After this step 4-aeC residues can be conjugated to biotinyl-n-hydroxysuccinimide ester yielding biotinylated DNA. This reaction allows the massive production of biotinylated probes. Labelled DNA can serve as molecular weight marker and positive control in Southern-blots. Moreover it can be useful in the study of DNA-protein interaction and in the isolation of d.s. DNA-binding proteins through chromatographic procedures.     

Cultivation of plant cells in a stirred vessel: effect of impeller design

Suspension cultures of Nicotiana tabacum were grown in a batch fermentor using different agitation systems. The effects of the impeller type, size, and agitation speed on the productivity of cell mass and secondary metabolites (phenolics) have been investigated. The use of a large, flat-bladed impeller (diameter 7.6 cm; width 14.0 cm) improved culture growth significantly over systems using a regular, flat-bladed impeller (diameter 5.6 cm; width 1.5 cm). An impeller of the same dimensions as the 14.0-cm-wide, large, flat-bladed impeller with sail cloth blades yielded a higher maximum growth rate in the exponential phase but resulted in a longer lag phase. Overall (intracellular and extracellular) phenolics concentration showed a direct relationship to culture growth rate whereas extracellular concentrations were a function of agitation conditions. Power consumption and flow pattern studies were also completed to further characterize the different impellers tested.     

The expression of granulocyte/macrophage colony-stimulating factor in activated mouse lymphocytes declines with age

The expression of granulocyte/macrophage colony-stimulating factor (GM-CSF) was studied in spleen lymphocytes isolated from male C57BL/6J mice of 6, 20, and 29 months of age. GM-CSF expression (biological activity and mRNA level) was maximum after culturing the lymphocytes for 45 hr with concanavalin A and phorbol myristate acetate. The induction of both GM-CSF activity and mRNA levels was observed to decline over 60% between 6 and 29 months of age. The age-related decline in the level of GM-CSF paralleled the age-related decline in the mRNA levels of interleukin-2 and interleukin-3.     

Expression of bacterial beta-galactosidase in animal cells

A recombinant plasmid containing the gene for bacterial β-galactosidase, situated close to the simian virus 40 early promoter, has been constructed. Transfection of CHO, L, and COS-1 cells with this plasmid led to the expression and appearance of the enzyme. Using this system, we have developed a series of promoter cloning vehicles capable of accepting promoter signals for animal genes.     

Changes in spatial distribution pattern during the larval stage of the fall webworm,Hyphantria cunea drury (Lepidoptera: Arctiidae)

Summary The changes in spatial distribution pattern during larval stage of the fall webworm,Hyphantria cunea were quantitatively investigated in the field experimental populations. The female adult deposits eggs as a cluster and the hatchlings make a compact colonial-web. In this period, the all-or-none type mortality which is characteristic in gregarious insect species was occasionary recognized before spinning a compact colonial-web. Once making a compact colonial-web, the larvae feed the leaves in the colonial-web up to about 5th instar. In this period, the movement of larvae occurred due to the local food shortage in a colonial-web and the expansion of colonial-web. As the larvae developed, the colonial-web was separated into several small groups. These larvae began to disperse about 5th instar. In this period, the local food shortage seems to be an important trigger for the larval dispersal. The mean concentration of larvae on leaves abruptly decreased, and finally the larvae became solitary at the 6th or 7th instars. The dispersal process in later larval stage is not necessarily due to the complete food shortage. The dispersal prior to the occurrence of food shortage may be a safety mechanism to protect the larvae from the food shortage.     

Galium procurrens,a new diploid relic species of theG. sylvaticum-group from the Balkan Peninsula

Galium procurrens is described as a new diploid relic species from Montenegro/N. Albania and SW. Bulgaria. It is related to the tetraploidG. laevigatum and other diploid and polyploid taxa of theG. sylvaticum-group inhabiting European deciduous forests.     

益生菌调控肠道屏障功能预防家禽球虫病的研究进展

球虫病给养禽业带来巨大经济损失,人们对绿色健康食品的迫切需求使球虫病的防控面临新的挑战.伴随世界"禁抗"进程的不断推进,家禽养殖业亟需一种安全有效的新型抗球虫方法.益生菌可竞争性排斥病原菌定殖以防止球虫病继发感染,可刺激宿主抗菌肽、黏蛋白和紧密连接蛋白的分泌以抵御球虫入侵,还可激活免疫反应以增强机体抗球虫感染的能力.本...     

有机物料对喀斯特地区石灰土有效N、Fe、Zn含量的影响

为研究不同有机物料对喀斯特石灰土元素有效性的影响,采用40 d的室内培养实验,比较了单独添加不同比例（1%、3%、5%）的生物质炭、鸡粪肥、羊粪肥对喀斯特石灰土有效N、Fe、Zn含量的影响。结果表明：添加生物质炭提高了喀斯特石灰土的pH值,而添加鸡粪肥和羊粪肥降低了喀斯特石灰土的pH值;添加3种有机物料均增加了喀斯特石灰土的有机质含量,大小关系为：生物质炭 > 鸡粪肥 > 羊粪肥,但添加鸡粪肥和羊粪肥土壤有机质的化学活性和微生物活性更高。受pH、有机质活性、碳氮比等因素的影响,添加鸡粪肥和羊粪肥能增加土壤有效N含量,但两种有机肥对土壤有效N的提高效果相差不大,而添加生物质炭反而降低了土壤有效N的含量;3种有机物料均能提高土壤的有效Fe和有效Zn含量,其中鸡粪肥效果最佳,其次为羊粪肥和生物质炭。当3种有机物料的添加比例为5%时,生物质炭处理土壤的有效N、Fe、Zn含量分别是对照处理的0.92、1.13、1.21倍;鸡粪肥处理土壤的有效N、Fe、Zn含量分别是对照处理的1.22、1.63和3.39倍;羊粪肥处理土壤的有效N、Fe、Zn含量分别是对照处理的1.27、1.34和2.59倍。所以,相对于生物质炭,有机粪肥对喀斯特地区的石灰土有更好的改良效果。