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31.
High-frequency disruption of the N-myc gene in embryonic stem and pre-B cell lines by homologous recombination. 总被引:15,自引:4,他引:11
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J Charron B A Malynn E J Robertson S P Goff F W Alt 《Molecular and cellular biology》1990,10(4):1799-1804
Identification of gene function has often relied on isolation of mutant cells in which expression of the gene was inactivated. Gene targeting by homologous recombination in tissue culture now may provide a technology to rapidly and directly produce such mutant mammalian cells. We demonstrate that selection of embryonic stem and pre-B cell lines for expression of a promoterless construct containing murine N-myc genomic sequences fused to a gene encoding neomycin resistance allows highly efficient recovery of variants in which the endogenous N-myc gene is disrupted. The high frequency of N-myc gene disruption by this method should permit targeted disruption of both allelic N-myc copies in various cell lines to study N-myc function. 相似文献
32.
A native high molecular complex (Mr 850000) containing about 50% of the allphycocyanin of the phycobilisome but lacking allophycocyanin B was separated from isolated phycobilisomes by gel electrophoresis. It was designated APCM since the large linker polypeptide LCM was exclusively localized in this complex. The complex exhibited a ?196°C fluorescence emission maximum at 673 nm (671 nm at 25°C). In addition, a core complex (designated APC, Mr≥1000000) consisting of both APCM and AP 680 was isolated by combined gel filtration and linear gradient centrifugation. At 25°C this complex showed dual emission peaks at 670 and 680 nm demonstrating functional independence of the terminal emitters. A complex similar to APCM can be isolated from phycobilisomes of Anabaena variabilis. This is evidence that APCM is the constitutive center of the tricylindrical core of hemidiscoidal cyanobacterial phycobilisomes. Two models summarizing the structural and functional consequences of the results are presented in the discussion. 相似文献
33.
Summary Two females showing partial expression of X-linked chondrodysplasia punctata were identified in a family. Bone dysplasia was caused by an aberrant X chromosome that had an inverse duplication of the segment Xp21.2–Xp22.2 and a deletion of Xp22.3-Xpter. To characterise the aberrant X chromosome, dosage blots were performed on genomic DNA from a carrier using a number of X-linked probes. Anonymous sequences from Xp21.2–Xp22.2 to which probes D2, 99.61, C7, pERT87-15, and 754 bind were duplicated on the aberrant X chromosome. The proposita was heterozygous for all these markers. Dosage blots also showed that the loci for steroid sulfatase and the cell surface antigen 12E7 (MIC2) were deleted as expected from the cytogenetic results. Mouse human cell hybrids were constructed that retained the normal X in the active state. Analysis of these hybrid clones for the markers from Xp21.2–Xp22.2 revealed that all the alleles of the informative markers, present in a single dosage in the genomic DNA, were carried on the normal X chromosome of the proposita. The duplicated X chromosome therefore had two identical alleles, indicating that the aberration resulted from an intrachromosomal rearrangement. 相似文献
34.
35.
Wolfgang Ludwig Michael Weizenegger Silvia Dorm Jan Andreesen Karl Heinz Schleifer 《FEMS microbiology letters》1990,71(1-2):139-143
A 1330 base-pair fragment of a 16S rRNA gene has been amplified, cloned and sequenced. Comparison to other 16S rRNA sequences of eubacteria showed that P. niger represents a deep branch within the subdivision "Gram-positive with Gram-negative cell walls". It is not related to peptostreptococci, representatives of this genus studied so far are more closely related to clostridia. 相似文献
36.
37.
Esmond J. Sanders Sara E. Zalik Wolfgang J. Schneider Irene M. Ledsham 《Development genes and evolution》1990,199(5):295-306
Summary Affinity purified preparations of the galactose-binding lectin from gastrulating chick blastoderms consist of three main polypeptides. Two of these have been identified as the 14 kD and 16 kD galactose-binding lectins. A third one migrates in SDS-PAGE gels with a relative molecular weight of 6,500±500 and has been identified as an apolipoprotein (Apo) of plasma very low density lipoproteins, Apo-VLDL-II. We have studied the localization of these polypeptides using immunofluorescence and ultrastructural immunocytochemistry with peroxidase and protein-A gold. The 14 kD lectin occurs in the intracellular yolk where it is mainly present within the electron lucent component. The 16 kD is also present in the intracellular yolk platelets, but tends to predominate in the electron-dense component. In addition, the 16 kD lectin is also present in pleiomorphic yolk-associated organelles and in the extracellular matrix. Apo-VLDL-II is also localized in the electron-lucent component of the yolk platelet and in the extracellular matrix. Our results suggest that the lectin(s) are associated with Apo-VLDL-II in the yolk platelet, and may subsequently become externalized. 相似文献
38.
Hans Georg B?umert Akitsugu Kenmoku Gert Middelhoff Franz Ortanderl Alexander Thrun Heinz Faulstich Wolfgang Schiebler Hugo Fasold 《Journal of Protein Chemistry》1988,7(5):571-580
With the aid of tartryl-bis--aminocaprylazide artificial dimers were produced from F actin from rabbit striated muscle. These derivatives will not polymerize by themselves but are able to copolymerize fully with native G actin. By modification of a single side chain per dimer, this copolymerization was completely inhibited. The dimers are able to activate subfragment I ATPase of myosin and bind to DNase I with inactivation of the enzyme in the same manner as native G actin. Within the dimer, one ADP is immobilized and will exchange against ATP extremely slowly. The dimers do not bind to the mushroom toxin phalloidin. 相似文献
39.
Dr. Wolfgang Schwabe Andreas Weihe Thomas Börner Manfred Henning Johannes-Günther Kohl 《Current microbiology》1988,17(3):133-137
The plasmid content and toxicity of nine different strains ofMicrocystis aeruginosa have been analyzed. The two toxic strains of the HUB Culture Collection were found to carry each two plasmids, pMA1 and pMA2, of 2.9 kb and 8.5 kb, respectively. In strains PCC 7813 and PCC 7820, also toxic, two different plasmids of 2.6 kb and 16 kb were detected. Hybridization experiments showed that there exists no sequence homology between the pMA plasmids and the plasmids found in the PCC strains; but the pMA plasmids hybridized to chromosomal DNA of the toxic strains PCC 7820, PCC 7813, HUB 063, and the nontoxic strain HUB 5-3. In nontoxic strains no or at most one plasmid of unstable occurrence could be detected. Only one of the toxic strains investigated, SAG 14.85 (NRC-1), contained no plasmid. 相似文献
40.
Wolfgang Walkowiak 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1988,164(1):31-41
Summary Male fire-bellied toads,Bombina bombina, are stimulated into calling by conspecific mating calls. They tend to synchronize their activity with other calling individuals, which leads to antiphonal calling. Behavioral physiological experiments revealed the existence of two different auditory filter systems which influence this acoustic behavior. One filter system is tuned to the recognition of conspecific mating calls, the other is part of a circuit that controls an acoustic jamming avoidance reaction. The two filter systems differ in several properties: filter bandwidth, threshold, and time constant. Neurophysiological recordings in the midbrain and the thalamus show that the filter characteristics can be explained by the coding properties of auditory neurons in these two brain nuclei. 相似文献