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1.
A simple new procedure was described for producing a sequential series of overlapping clones for use in DNA sequencing. The technique used single-stranded M13 DNA and complementary DNA oligomers to form specific cleavage and ligation substrates. It was, therefore, independent of the sequence of the DNA cloned into the vector. Deletions of varying sizes were generated from one end of the insert through the 3' to 5' exonuclease activity of T4 DNA polymerase. The approximate size of the deletion and therefore the starting point for DNA sequencing could be estimated by electrophoresis of the subcloned phage DNA on a agarose gel. This greatly reduced the number of templates that must be sequenced to obtain a complete sequence. The entire procedure could be carried out in one tube in less than a day. The procedure was used to subclone and sequence the maize mitochondrial 18 S rDNA and 5' flanking region (2622 bases) in less than a week. Other applications of oligomers and single-stranded DNA in the construction of insertions, deletions, and cDNAs are discussed. 相似文献
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Ana Isabell Paulino Aud Larsen Gunnar Bratbak Dale Evens Svein Rune Erga Einar Bye-Ingebrigtsen 《Marine Biology Research》2018,14(5):421-435
Changes in phytoplankton community composition potentially affect the entire marine food web. Because of seasonal cycles and inter-annual variations in species composition, long-term monitoring, covering many sequential years, is required to establish a baseline study and to reveal long-term trends. The current study describes the phytoplankton biomass variations and species composition in relation to hydrographic and meteorological conditions in the Raunefjord, western Norway, over a 6-year period from 2001 to 2006. The extent of inflow or upwelling in the fjord varied from year to year and resulted in pronounced differences in water column stability. The annual phytoplankton community succession showed some repeated seasonal patterns, but also high variability between years. Two to four diatom blooms were observed per year, and the spring blooms occurring before water column stratification in March were dominated by Skeletonema marinoi and Chaetoceros socialis, and other Chaetoceros and Thalassiosira spp. Blooms of the haptophytes Phaeocystis pouchetii and Emiliania huxleyi were irregular and in some years totally absent. Although E. huxleyi was present all year round it appeared in bloom concentrations only in 2003, when the summer was warm and the water column characterized by high surface temperatures and pronounced stratification. The annual average abundance of both diatoms and flagellates increased during the six years. Despite the high variation from year to year, our investigation provides valuable knowledge about annual phytoplankton community patterns in the region, and can be used as a reference to detect possible future changes. 相似文献
4.
We have analysed the lineage of olfactory receptor neurons usinga replication-incompetent retrovirus injected beneath the olfactoryepithelium of young rats. There are two major types of clustersof infected cells seen at 540 days after infection: (i)horizontal basal cells (HBCs); (ii) variable numbers of globosebasal cells (GBCs), and immature and mature sensory neurons.Olfactory nerve lesion increased the frequency of the globose/sensoryneuron clusters, as well as the number of cells/cluster, butdid not change the number of HBC clusters or cells/cluster.No clusters contained sustentacular cells. These data indicatethat, at least in young rats: (i) HBCs are not precursors ofolfactory neurons; (ii) there is a lineage path from GBCs tomature neurons; and (iii) sustentacular cells arise from a separatelineage. 相似文献
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Experiments are described which were designed to investigate the significance of individual differences in aggressiveness between polecats and the relation between the familiarity of the opponent and the pattern and outcome of fighting.
The behaviour of Mustela putorius, M. furo and hybrids between the two species was investigated in an indoor arena of 16 m2 .
Two kinds of fighting between male polecats were recorded, "companion fighting" between cage mates, in which biting was inhibited and neither of the opponents became intimidated and "uninhibited fighting" between unfamiliar individuals from which a winner and loser generally emerged and a rank order formed.
Individual differences in aggressiveness were assessed by means of a scoring system which statistical analysis showed, generally, to be consistent for any one individual. The rank of an animal, however, was found not to be invariably related to its aggression score.
A number of other factors influencing fighting were investigated, the earlier introduction of one individual into the arena increases its chances of winning a fight; when offered a choice, male polecats fight strangers in preference to cage mates; and the separation of a group of cage mates for as little as 48 hours induces them to behave towards one another like strangers with the result that a rank order is formed. Factors which had no apparent effect on fighting between males were the weight of the individual or the presence of females.
The attributes of familiar and unfamiliar opponents are discussed in the light of these findings. 相似文献
The behaviour of Mustela putorius, M. furo and hybrids between the two species was investigated in an indoor arena of 16 m
Two kinds of fighting between male polecats were recorded, "companion fighting" between cage mates, in which biting was inhibited and neither of the opponents became intimidated and "uninhibited fighting" between unfamiliar individuals from which a winner and loser generally emerged and a rank order formed.
Individual differences in aggressiveness were assessed by means of a scoring system which statistical analysis showed, generally, to be consistent for any one individual. The rank of an animal, however, was found not to be invariably related to its aggression score.
A number of other factors influencing fighting were investigated, the earlier introduction of one individual into the arena increases its chances of winning a fight; when offered a choice, male polecats fight strangers in preference to cage mates; and the separation of a group of cage mates for as little as 48 hours induces them to behave towards one another like strangers with the result that a rank order is formed. Factors which had no apparent effect on fighting between males were the weight of the individual or the presence of females.
The attributes of familiar and unfamiliar opponents are discussed in the light of these findings. 相似文献
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Nisin stimulates oxygen consumption by Staphylococcus aureus and Escherichia coli. 总被引:2,自引:2,他引:0
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Nisin stimulated oxygen consumption by nongrowing, glucose-metabolizing Staphylococcus aureus and Escherichia coli cells, indicating a protonophore mode of action. A similar stimulation in E. coli cells osmotically stressed to disrupt the outer cell membrane confirmed the cytoplasmic membrane as the site of nisin action and showed that nisin uptake was not prevented by the outer membrane. 相似文献
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Silvia Penuela Alexander W Lohman Wesley Lai Laszlo Gyenis David W Litchfield Brant E Isakson Dale W Laird 《Channels (Austin, Tex.)》2014,8(2):124-130
The pannexin family of channel-forming proteins is composed of 3 distinct but related members called Panx1, Panx2, and Panx3. Pannexins have been implicated in many physiological processes as well as pathological conditions, primarily through their function as ATP release channels. However, it is currently unclear if all pannexins are subject to similar or different post-translational modifications as most studies have focused primarily on Panx1. Using in vitro biochemical assays performed on ectopically expressed pannexins in HEK-293T cells, we confirmed that all 3 pannexins are N-glycosylated to different degrees, but they are not modified by sialylation or O-linked glycosylation in a manner that changes their apparent molecular weight. Using cell-free caspase assays, we also discovered that similar to Panx1, the C-terminus of Panx2 is a substrate for caspase cleavage. Panx3, on the other hand, is not subject to caspase digestion but an in vitro biotin switch assay revealed that it was S-nitrosylated by nitric oxide donors. Taken together, our findings uncover novel and diverse pannexin post-translational modifications suggesting that they may be differentially regulated for distinct or overlapping cellular and physiological functions. 相似文献