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1.
Polymorphism of animal mitochondrial DNA (mtDNA) has been shown to involve
point mutations and limited length variations affecting essentially
noncoding regions. In two wild mice of the European subspecies Mus mus
musculus we found a mitochondrial mutant with a very large deletion in a
coding region. The deletion is 5 kbp long (31% of the mitochondrial
chromosome) and encompasses six tRNA genes and seven protein genes. The two
mice were heteroplasmic: they contained a mixture of normal mtDNA and the
deletion mutant. Although the latter is functionally defective, it
represents 78%-79% of the mtDNA molecules in our preparations from each
animal.
相似文献
2.
A technique involving culture in soft agar was used for the assay of forward mutation of V79 cells to 6-thioguanine (6TG) resistance. The main reason for the use of soft agar was to prevent reduction in recovery of mutants depending on the cell density plated for mutation selection, which is the chief problem in the liquid method, and which results mainly from metabolic co-operation due to cell-to-cell contact.V79 cells grew well in fortified soft agar medium (DMEM + 20% FBS) showing cloning efficiencies (>80%) as high as in liquid culture. Therefore, V79/HGPRT mutagenesis could be assayed quantitatively in soft agar culture.The frequency of 6TG-resistant colonies in agar selective medium increased linearly with increase in concentration of EMS. Toxicity and mutagenic responses were greater in soft agar than in liquid culture.In cultures of untreated and EMS-treated cells, more than 95% of the 6TG-resistant colonies isolated were aminopterin-sensitive.Use of soft agar for selection prevented the reduction in the number of mutants with increase in the size of incula on plating up to 1?2 × 106 cells per 9-cm dish: in liquid culture, even with a lower plating number (2 × 105 cells per 9-cm dish), a notable reduction in numbers of mutants was observed. This character was re-examined in a reconstruction experiment. The results show that, when up to 2 × 106 cells were plated per 9-cm dish, 6TG-resistant cells were almost completely recovered from the soft agar medium, whereas only 10% were recovered from liquid culture. 相似文献
3.
Makiko Kawagishi-Kobayashi Masayuki Yamamoto Akira Ishihama 《Molecular & general genetics : MGG》1996,250(1):1-6
Local sequence similarity exists between the subunit 2 of eukaryotic RNA polymerases II and the barnase-type bacterial RNases. The RNase-like domain from the Rpb2 ofSchizosaccharomyces pombe was expressed inEscherichia coli as a GST fusion protein and examined for its RNase activity. When the GST fusion protein was incubated in vitro with32P-labeled RNA, the RNA degradation activity was less than 0.1%, if any, of the level of synthetic barnase. In order to check the in vivo function of this region, we constructed two mutantrpb2 alleles,rpb2
E357A
andrpb2
H3a6L
, each carrying a single amino acid substitution at the site correponding to one of the three essential amino acid residues forming the catalytic site in barnase (mutation of barnase at the corresponding sites results in complete loss of RNase activity) and five other mutantrpb2 alleles, each carrying a single mutation at various positions within the RNase-like domain but outside the putative catalytic site for RNase activity. When these mutantrpb2 alleles were expressed in anrpb2-disruptedS. pombe strain, all the mutants grew as well as the wild-type parent and did not show any clear defective phenotypes. These results suggest either that the RNase-like domain in Rpb2 does not function as an RNase in vivo or that the RNase activity of this domain, if present at all, is not essential for cell growth. 相似文献
4.
Lentinan, a -1,6;1,3-glucan, is tumor-specific for transplantable mouse solid-type tumors and it also stimulates the production of acute phase proteins (APPs). The APP response to lentinan is of the delayed type (DT-APR) and differs from that to lipopolysaccharide, which is acute. We found that the responses were genetically controlled in mice and that low responsiveness is dominant (Maeda et al. 1991). Using 123 segregants of crosses between SWR/J (a high responder) andMus spretus (a low responder), we analyzed the linkage between DT-APR responsiveness and the DNA polymerase chain reaction-simple sequence lenght polymorphism (PCR-SSLP) phenotype using 80 chromosome-specific microsatellite markers. We identified two loci (ltn1.1 andltn1.2) responsible for DT-APR.ltn1.1 is closely linked toD3Mit11 on chromosome 3 andltn1.2 toD11Nds9 on chromosome 11 (P<0.001). The linkage analysis also suggested thatltn1.2 is the major determinant for DT-APR. Correlation between lentinan-specific IL-6 mRNA expression (the late expression) controlled recessively and DT-APR induction suggests that theltn1 loci control some process(es) of IL-6 expression in the regulation step before NF-IL6. 相似文献
5.
Mitochondrial DNA polymorphisms in subterranean mole-rats of the Spalax ehrenbergi superspecies in Israel, and its peripheral isolates 总被引:1,自引:0,他引:1
Nevo E; Honeycutt RL; Yonekawa H; Nelson K; Hanzawa N 《Molecular biology and evolution》1993,10(3):590-604
Patterns of mitochondrial DNA (mtDNA) variation were examined in 133
mole-rats constituting all four chromosomal species (2n = 52, 2n = 54, 2n =
58, and 2n = 60) of the Spalax ehrenbergi superspecies in Israel, as well
as the peripheral isolates of 2n = 60. In the main range of the complex, a
total of 28 mtDNA haplotypes were found in 64 mole-rats, with most
haplotypes being unique to either a single chromosomal species or
population. mtDNA divergence increased from low to high diploid number in a
north-to-south direction in Israel. Overall levels of mtDNA diversity were
unexpectedly the highest in the 2n = 60, the youngest species of the
complex. The mtDNA haplotypes can be separated into two major groups, 2n =
52-54 and 2n = 58-60, and a phylogenetic analysis for each group revealed
evidence of a few haplotypes not sorted by diploid number. The overall
patterns of mtDNA divergence seen within and among the four chromosomal
species are consistent with the parapatric mode of speciation as suggested
from previous studies of allozyme and DNA hybridization. In a separate data
set the patterns of mtDNA variation were examined across the main
geographic range and across peripheral semi-isolates and isolates of the 2n
= 60 chromosomal species. Fifteen haplotypes were found in 69 mole-rats.
High levels of mtDNA diversity characterized the main range, semi-isolated,
and even some desert isolated populations. The peripheral isolates contain
much mtDNA diversity, including novel haplotypes.
相似文献
6.
Tamio Ohno Jun-ichi Katoh Yoshiaki Kikkawa Hiromichi Yonekawa Masahiko Nishimura 《Experimental Animals》2003,52(5):415-417
To develop SMXA recombinant inbred (RI) strains as more valuable genetic resources, 302 microsatellite (Mit) loci were added to the strain distribution patterns (SDP) reported previously. The improved SDP were constructed in a total of 1085 loci containing 484 Mit markers, 571 restriction landmark genomic scanning (RLGS) spot markers and 30 others. This substantially improved SDP can be freely accessed on our homepage (http://www.med.nagoya-u.ac.jp/sisetu/SDP.htm). 相似文献
7.
We have developed a new method for the prediction of the lateral and the rotational positioning of transmembrane helices, based upon the present status of knowledge about the dominant interaction of the tertiary structure formation. The basic assumption about the interaction is that the interhelix binding is due to the polar interactions and that very short extramembrane loop segments restrict the relative position of the helices. Another assumption is made for the simplification of the prediction that a helix may be regarded as a continuum rod having polar interaction fields around it. The polar interaction field is calculated by a probe helix method, using a copolymer of serine and alanine as probe helices. The lateral position of helices is determined by the strength of the interhelix binding estimated from the polar interaction field together with the length of linking loop segments. The rotational positioning is determined by the polar interaction field, assuming the optimum lateral configuration. The structural change due to the binding of a prosthetic group is calculated, fixing the rotational freedom of a helix that is connected to the prosthetic group. Applying this method to bacteriorhodopsin, the optimum lateral and rotational positioning of transmembrane helices that are very similar to the experimental configuration was obtained. This method was implemented by a software system, which was developed for this work, and automatic calculation became possible for membrane proteins comprised of several transmembrane helices. © 1995 Wiley-Liss, Inc. 相似文献
8.
Using cDNA of the human poliovirus receptor (PVR) as a probe, two types of cDNA clones of the monkey homologs were isolated from a cDNA library prepared from an African green monkey kidney cell line. Either type of cDNA clone rendered mouse L cells permissive for poliovirus infection. Homologies of the amino acid sequences deduced from these cDNA sequences with that of human PVR were 90.2 and 86.4%, respectively. These two monkey PVRs were found to be encoded in two different loci of the genome. Evolutionary analysis suggested that duplication of the PVR gene in the monkey genome had occurred after the species differentiation between humans and monkeys. The NH2-terminal immunoglobulin-like domain, domain 1, of the second monkey PVR, which lacks a putative N-glycosylation site, mediated poliovirus infection. In addition, a human PVR mutant without N-glycosylation sites in domain 1 also promoted viral infection. These results suggest that domain 1 of the monkey receptor also harbors the binding site for poliovirus and that sugar moieties possibly attached to this domain of human PVR are dispensable for the virus-receptor interaction. 相似文献
9.
H Yonekawa K Moriwaki O Gotoh N Miyashita S Migita F Bonhomme J P Hjorth M L Petras Y Tagashira 《Differentiation; research in biological diversity》1982,22(3):222-226
To determine the origins of laboratory mice, the restriction patterns of mitochondrial DNAs (mtDNAs) from various strains were compared with those of relevant subspecies and/or races of Mus musculus. In most strains and substrains of laboratory mice examined (50/55), the cleavage patterns were identical to those of the European subspecies M. m. domesticus. Those that varied include two sublines of NZB, the strain NZC, and the Japanese strain RR. The NZB and NZC patterns were identical to that of the European subspecies M. m. brevirostris, which itself has restriction patterns similar to M. m. domesticus. On the other hand, the RR pattern was identical to M. m. molossinus-like mice trapped in Western China and slightly different from Japanese M. m. molossinus. These findings suggest that the strains NZB and NZC stemmed from a European founder stock which differed from the ancestral stocks of other laboratory strains and that the ancestral mice of the RR strain had been transported from China to Japan. Therefore, most laboratory strains of mice are derived from the European subspecies M. m. domesticus while M. m. brevirostris and M. m. molossinus have made minor contributions. M. m. musculus does not appear to have made any contribution. 相似文献
10.
Mitsumasa Haruna Makiko Kato Kazuo Ito Toshiaki Nikai Hisayoshi Sugihara Hiroyuki Muratat 《Phytochemistry》1981,20(11):2583-2584
Angeloylcumambrin-B, a new antimicrobial guaianolide sesquiterpene lactone, was isolated from Chrysanthemum ornatum and the structure was determined by a combination of chemical and physical methods. 相似文献