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A composite mouse androgen receptor DNA sequence was obtained by amplifying genomic DNA or cDNA using the polymerase chain reaction. The open reading frame was 2,697 basepairs, encoding a polypeptide of 899 amino acids (98,204 mol wt). Amino acid sequence comparisons indicated that the mouse androgen receptor (AR) is 97% homologous with rat AR and 83% with human AR. The amino acid sequences of the three receptors are identical within the DNA- and steroid-binding domains. Northern blot analysis revealed the predominant mouse AR mRNA to be 10 kilobases (kb). A 1.7-kb mRNA species was detected in mouse kidney using a cDNA probe containing only 5' untranslated AR sequence. Lack of hybridization with AR-coding sequence probes suggested that the 1.7-kb mRNA was not a truncated form of AR mRNA. Sequencing of genomic DNA isolated from testicular feminized (Tfm) mice revealed a single base deletion in the N-terminal domain, resulting in a frameshift mutation. Cycloheximide treatment caused a dramatic increase in AR mRNA in kidneys of Tfm mice, but not wild-type mice, suggesting that the Tfm mutation results in an unstable AR mRNA.  相似文献   

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The content of epidermal growth factor (EGF) as a high molecular weight complex (HMW-EGF) in the submandibular glands of mice was measured simply by a single radial immunodiffusion method. In female mice, the amount of HMW-EGF was increased 10-fold by tri-iodo-L-thyronine (T3) and 60-fold by 5 alpha-dihydrotestosterone (5 alpha-DHT). In mice with testicular feminization (Tfm), which are genetically deficient in androgen receptor, T3 but not 5 alpha-DHT increased the HMW-EGF from a non-detectable level to 5.4 +/- 0.94 micrograms/mg protein. It was concluded that EGF is also synthesized under the control of thyroid hormone in vivo, and that androgen was not involved in this induction of EGF by thyroid hormone.  相似文献   

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Five independent clones containing the natural chicken ovomucoid gene have been isolated from a chicken gene library. One of these clones, CL21, contains the complete ovomucoid gene and includes more than 3 kb of DNA sequences flanking both termini of the gene. Restriction endonuclease mapping, electron microscopy and direct DNA sequencing analyses of this clone have revealed that the ovomucoid gene is 5.6 kb long and codes for a messenger RNA of 821 nucleotides. The structural gene sequence coding Ifor the mature messenger RNA is split into at least eight segments by a minimum of seven intervening sequences of various sizes. The shortest structural gene segment is only 20 nucleotides long. All seven intervening sequences are located within the peptide coding region of the gene, and the sequences at the 5' and 3' untranslated regions of the mRNA are not interrupted by intervening sequences. The DNA sequences of the regions flanking the 5' and 3' termini of the gene have been determined. Thirty nucleotides before the start of the messenger RNA coding sequence is the heptanucleotide TATATAT, which is also present in a similar location relative to the chicken ovalbumin gene and other unique sequence eucaryotic genes. This sequence resembles that of the Pribnow box in procaryotic genes where a promoter function has been implicated. Seven nucleotides past the 3' end of the gene is the tetranucleotide TTGT, a sequence found to be present at identical locations as either TTTT or TTGT in other eucaryotic genes that have been sequenced. These conserved DNA sequences flanking eucaryotic genes may serve some regulator function in the expression of these genes.  相似文献   

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Androgen regulation of MAK mRNAs in mouse kidney   总被引:5,自引:0,他引:5  
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Summary The X-linked testicular feminization mutation (Tfm) of the mouse leads to androgen insensitivity of target cells. Through the autosomal sex reversed (Sxr) factor we have converted female heterozygotes into males. Due to X-inactivation, mosaic animals arise which are composed of androgen sensitive wild-type and androgen insensitive Tfm cells. In the androgen dependent striated urethral muscle, Tfm and wild-type cells fuse and form multinucleated muscle fibres. In the muscle fibres the Tfm nuclei are exposed to the intact cytoplasmic testosterone receptor complex coded for by the wildtype nuclei. We ask the question whether under these conditions RNA synthesis can be stimulated in the Tfm nuclei. Castrated mosaic animals were injected with testosterone, and incorporation of 3H-uridine was studied by autoradiography. We found two classes of muscle cell nuclei, those with low grain counts corresponding to the Tfm controls and those with high grain counts corresponding to the stimulated male controls. The results indicate that the Tfm nuclei are not stimulated by the intact testosterone receptor complex.This study is dedicated to Prof. Dr. W. Graumann on occasion of his 65th birthday  相似文献   

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Reduced reproductive performance in androgen-resistant Tfm/Tfm female mice   总被引:1,自引:0,他引:1  
Androgen-resistant female mice (Tfm/Tfm) homozygous for the mutant gene Tfm were bred by making use of males chimaeric for the Tfm gene. All seven Tfm/Tfm females found were fertile, confirming that a normal level of androgen receptor protein is not essential for reproduction in female mice. However, when five of the seven were studied throughout their reproductive life they proved to have impaired reproductive performance and premature cessation of reproduction. No impairment of reproduction was seen in heterozygous Tfm/+ females. The ovarian histology suggested that in Tfm/Tfm the normal ageing processes were accelerated. This work is consistent with the work of others in that androgen is involved in the control of follicular maturation and atresia, and that the effect is mediated by the androgen receptor coded by the Tfm locus.  相似文献   

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We have detected a mouse mammary tumor virus (MMTV)-specific 1.7-kilobase (kb) polyadenylated RNA in mammary glands of several mouse strains. In BALB/c mice, it is the only MMTV-specific RNA species present. C3H and GR mammary glands and tumors contain, in addition, 3.8- and 7.8-kb MMTV RNAs. Nuclease S1 analysis was performed to map 1.7-kb polyadenylated RNA. It contains predominantly long terminal repeat (LTR) sequences. The 5' end maps approximately 134 nucleotides upstream from the 3' end of the LTR. Colinearity with complete proviral DNA continues to a site about 153 nucleotides downstream from the left (5') LTR. No sequences from the middle part of proviral DNA were found. Colinearity with proviral DNA is resumed 72 nucleotides upstream from the right (3') LTR. The nucleotide sequence in this area is TTCCAGT, which is a splice acceptor consensus sequence. The anatomy of 1.7-kb RNA indicates that it may serve as a messenger for the 36,700-dalton protein encoded by the LTRs of MMTV.  相似文献   

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The role of androgens on the sexual dimorphism of mandible shape was investigated in mice carrying the X-linked gene for testicular feminization (Tfm), which is known to determine a profound insensitivity to testosterone and is associated with a severe reduction in androgen receptor levels in Tfm/Y males. Mandible shape analysis in an inbred strain of mice segregating for the Ta (tabby) and Tfm mutations showed that the sexual dimorphism observed between +Ta/+Ta females and +Ta/Y males almost disappeared between Tfm+/+Ta females and Tfm+/Y males. In addition, a canonical discriminant analysis showed that these two closely related classes, Tfm+/+Ta and Tfm+/Y, are readily differentiated from both the +Ta/+Ta and +Ta/Y classes. These results suggest that androgens are involved in the mandible shape sexual dimorphism and play a role in mandibular development in both males and females.  相似文献   

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Sequence organization of feline leukemia virus DNA in infected cells   总被引:15,自引:0,他引:15       下载免费PDF全文
A restriction site map has been deduced of unintegrated and integrated FeLV viral DNA found in human RD cells after experimental infection with the Gardner-Arnstein strain of FeLV. Restriction fragments were ordered by single and double enzyme digests followed by Southern transfer (1) and hybridization with 32P-labeled viral cDNA probes. The restriction map was oriented with respect to the 5' and 3' ends of viral RNA by using a 3' specific hybridization probe. The major form of unintegrated viral DNA found was a 8.7 kb linear DNA molecule bearing a 450 bp direct long terminal redundancy (LTR) derived from both 5' and 3' viral RNA sequences. Minor, circular forms, 8.7 kb and 8.2 kb in length were also detected, the larger one probably containing two adjacent copies of the LTR and the smaller one containing one comtaining one copy of the LTR. Integrated copies of FeLV are colinear with the unintegrated linear form and contain the KpnI and SmaI sites found in each LTR.  相似文献   

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Studies of cloned sequences from four Drosophila heat shock loci.   总被引:45,自引:0,他引:45  
R Holmgren  K Livak  R Morimoto  R Freund  M Meselson 《Cell》1979,18(4):1359-1370
DNA cloned from the D. melanogaster (Oregon R) heat shock loci at 63BC and 95D codes for the 83,000 and the 68,000 dalton heat shock proteins, respectively. Both coding sequences occur once per haploid genome. Sequences complementary to messenger RNA for the 70,000 dalton heat shock protein are represented five times, twice at 87A and three times at 87 C. The copies at 87A differ characteristically from those at 87C in an interval of a few hundred bp near the 5' end of the messenger sequence, and the corresponding two classes of hsp 70 messenger RNA are found on polysomes after heat shock. Within this differential region, there is about 15% divergence between messenger sequences cloned from the two loci, while in the rest of the messenger region examined the homology is much closer although still imperfect. Unexpectedly, considerable homology is found between the sequence for the 68,000 dalton heat shock protein at 95D and the sequences for the 70,000 dalton protein at 87A and 87C, and between these sequences and a site in 87D. Messenger RNA molecules of 2.4, 2.55 and 3.05 kb code for the 68,000, 70,000 and 83,000 dalton heat shock proteins and hybridize to apparently uninterrupted DNA sequences of 2.1, 2.25 and 2.6 kb, respectively.  相似文献   

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Fractionation of messenger activities isolated from the cytoplasm of HeLa cells late in infection with adenovirus type 2 reveals that viral polypeptides III and pVII are each synthesized from two different-sized mRNA's. the major messenger activity for each protein has the same sedimentation rate as that previously reported by Anderson et al. (Proc. Natl. Acad. Sci. U.S.A. 71:2756-2760, 1974). The minor messenger activities for III and pVII sediment more rapidly and are not aggregates of the major mRNA's for these proteins. The two minor messenger activities cosediment with two polyadenylated RNA species which are labeled late in infection with 32P and whose molecular weights are estimated to be 2.9 x 10(6) and 2.4 x 10(6). Both of these species hybridize to adenovirus type 2 DNA specific for the mRNA family that is 3' coterminal at adenovirus type 2 map position 49.5 and the mRNA family that is 3' coterminal at 62.0. This is consistent with the possibility that these RNAs have 5'-terminal sequences identical to those of the normal mRNA's for III and pVII but are 3' coterminal at map position 62, the normal 3' terminus of the mRNA's for polypeptides II and pVI. These species are not found in polyadenylated RNA isolated from the nucleus, suggesting that the minor mRNA species are cytoplasmic RNAs.  相似文献   

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M Reitman  E Lee    H Westphal 《Nucleic acids research》1995,23(10):1790-1794
We have shown previously that the chicken beta A-globin gene, with its 3' enhancer, is expressed in a copy number-dependent manner in transgenic mice. The expression level was low but increased approximately 6-fold upon inclusion of 11 kb of upstream DNA containing four DNase I hypersensitive sites. To study the effect of the individual upstream hypersensitive sites on transgene expression, we produced lines of mice in which the individual upstream sites were linked to the beta A gene and enhancer. RNA levels were measured in blood from adult animals. With each of these four constructs, the level of transgene RNA per DNA copy varied over a > 20-fold range. These data suggest that addition of a hypersensitive site to the beta A-globin/enhancer region abrogates its position independent expression. The average beta A-globin expression per copy in the lines carrying an upstream site was comparable with that in lines without an upstream site. Thus, no single upstream hypersensitive site accounts for the higher level of beta A-globin expression seen in mice containing the complete upstream region. We had shown previously that control of the chicken beta-globin cluster is distributed between at least two regions, the beta A/epsilon enhancer and the upstream region. Our current results suggest that the control mediated by the upstream DNA is itself distributed and is not due to a single hypersensitive site.  相似文献   

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