Mapping of testis-determining locus on Yp by the molecular genetic analysis of XX males and XY females

Sex reversal in males with female karyotypes is likely to be caused by the presence of cytogenetically undetectable Y-chromosomal DNA sequences that include the testis-determining gene(s). Studying a total of sixteen 46,XX males and one 47,XXX male, we detected Y-chromosomal DNA in 13 of the XX males (i.e. 80 %) and in the 47,XXX male. The amount of Y-chromosomal DNA present in the patients varied between individuals. This allowed the construction of a molecular map of the Y-chromosome short arm. The putative testis-determining locus was assigned to the more distal portion of Yp, yet proximal to the pseudoautosomal region. Mapping of the testis-determining locus was complemented by molecular findings in 46,XY females. These individuals may carry microdeletions of the portion of Yp that appears to be required for normal male gonadogenesis. The deletions detected in 46,XY females always included those Y-chromosomal DNA sequences that were found in most 46,XX males. Furthermore, the same DNA sequences were missing in a female with a 46,X,dic(Y) karyotype. The observations suggest that some of our DNA probes hybridize with Y-chromosomal DNA sequences within a few million base pairs of the testis locus. Chromosome walking and pulscd-field gel electrophoresis investigations have been initiated in order to isolate those Y-chromosomal DNA sequences that are required for normal testicular development.

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Identification of Gene Related to Hard Bunch Phenotype in Oil Palm (Elaeis guineensis Jacq.)

Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) ◽

10.24831/jai.v43i2.10421 ◽

2015 ◽

Vol 43 (2) ◽

pp. 147

Author(s):

Roberdi , ◽

Sobir , ◽

Sudirman Yahya ◽

Nurita Toruan-Mathius ◽

Tony Liwang

Keyword(s):

Dna Sequences ◽

Oil Palm ◽

Elaeis Guineensis ◽

Molecular Genetic ◽

Molecular Genetic Analysis ◽

Target Sequence ◽

Polymorphic Markers ◽

Copia Retrotransposon ◽

Selection Of

ABSTRACTMolecular genetic analysis of hard bunch phenomenon in oil palm was done in order to elucidate the role of genetic factor underlying hard bunch in oil palm plantation. The aim of this study was to identify the AFLP primer combination that co-segregates with hard bunch phenotype related gene in oil palm. Molecular analysis was done by bulk segregant analysis approach. DNA was isolated from leaves of the normal and hard bunch palm. DNA from ten individual palms from each category were pooled and used as a template. A total of 56 AFLP primer combinations were selected for selection of polymorphic primer, and as a result it was found that 22 AFLP primer combinations (39.28%) were polymorphic. A total of 48 individual of palm DNA containing 24 individual for each group were further genotyped by those 22 polymorphic markers. Of these, one AFLP primer combination (E-ACC/M-CTG) was obtained as a co-segregated marker that distinguished the hard bunch DNA from the normal one. Based on the analysis of the target sequence aligned to the oil palm DNA sequences available in database, we found that our sequence has similarity with Ty-1 copia retrotransposon. This sequence distribute in all 16 linkage group of oil palm genome.Keywords: abnormal fruits, AFLP, oil palm, Ty-1 copia retrotransposon

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A delta-globin gene derived from patients with homozygous delta zero- thalassemia functions normally on transient expression in heterologous cells

Blood ◽

10.1182/blood.v70.3.809.bloodjournal703809 ◽

1987 ◽

Vol 70 (3) ◽

pp. 809-813

Author(s):

T Nakamura ◽

Y Takihara ◽

Y Ohta ◽

S Fujita ◽

Y Takagi ◽

...

Keyword(s):

Globin Gene ◽

Molecular Genetic ◽

Regulation Of Gene Expression ◽

Molecular Genetic Analysis ◽

Nucleotide Sequence Analysis ◽

Complete Suppression ◽

Erythroid Cells ◽

Base Pairs ◽

Chain Synthesis ◽

A Chain

Three Japanese individuals with homozygous delta zero-thalassemia from different families were the subjects of molecular genetic analysis. They were homozygous for seven polymorphic sites in the beta-globin gene cluster. Nucleotide sequence analysis of the delta-globin gene cloned from each patient revealed a single nucleotide substitution (T- C) 77 base pairs 5′ to the cap site, just upstream of the CCAAC box of the delta-globin gene. When introduced into COS cells, the gene was expressed at normal levels with proper processing of RNA. These results suggest that the complete suppression of delta-globin chain synthesis in these patients is not due to a defective promoter, a defective RNA processing or a chain terminator mutation, but rather to impaired regulation of gene expression specific to erythroid cells. The region around the CCAAC box may have a significant role in expression of the delta-globin gene in erythroid cells.

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A delta-globin gene derived from patients with homozygous delta zero- thalassemia functions normally on transient expression in heterologous cells

Blood ◽

10.1182/blood.v70.3.809.809 ◽

1987 ◽

Vol 70 (3) ◽

pp. 809-813 ◽

Cited By ~ 15

Author(s):

T Nakamura ◽

Y Takihara ◽

Y Ohta ◽

S Fujita ◽

Y Takagi ◽

...

Keyword(s):

Globin Gene ◽

Molecular Genetic ◽

Regulation Of Gene Expression ◽

Molecular Genetic Analysis ◽

Nucleotide Sequence Analysis ◽

Complete Suppression ◽

Erythroid Cells ◽

Base Pairs ◽

Chain Synthesis ◽

A Chain

Abstract Three Japanese individuals with homozygous delta zero-thalassemia from different families were the subjects of molecular genetic analysis. They were homozygous for seven polymorphic sites in the beta-globin gene cluster. Nucleotide sequence analysis of the delta-globin gene cloned from each patient revealed a single nucleotide substitution (T- C) 77 base pairs 5′ to the cap site, just upstream of the CCAAC box of the delta-globin gene. When introduced into COS cells, the gene was expressed at normal levels with proper processing of RNA. These results suggest that the complete suppression of delta-globin chain synthesis in these patients is not due to a defective promoter, a defective RNA processing or a chain terminator mutation, but rather to impaired regulation of gene expression specific to erythroid cells. The region around the CCAAC box may have a significant role in expression of the delta-globin gene in erythroid cells.

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Construction, replication, and chromatin structure of TRP1 RI circle, a multiple-copy synthetic plasmid derived from Saccharomyces cerevisiae chromosomal DNA.

Molecular and Cellular Biology ◽

10.1128/mcb.2.3.221 ◽

1982 ◽

Vol 2 (3) ◽

pp. 221-232 ◽

Cited By ~ 88

Author(s):

V A Zakian ◽

J F Scott

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Replication ◽

Dna Sequences ◽

Recombinant Dna ◽

Suitable Model ◽

Chromosomal Dna ◽

Base Pairs ◽

Autonomously Replicating Sequence ◽

Cell Cycles ◽

Initiation Of Replication

Transformation studies with Saccharomyces cerevisiae (bakers' yeast) have identified DNA sequences which permit extrachromosomal maintenance of recombinant DNA plasmids in transformed cells. It has been hypothesized that such sequences (called ARS for autonomously replicating sequence) serve as initiation sites for DNA replication in recombinant DNA plasmids and that they represent the normal sites for initiation of replication in yeast chromosomal DNA. We have constructed a novel plasmid called TRP1 R1 Circle which consists solely of 1,453 base pairs of yeast chromosomal DNA. TRP1 RI Circle contains both the TRP1 gene and a sequence called ARS1. This plasmid is found in 100 to 200 copies per cell and is relatively stable during both mitotic and meiotic cell cycles. Replication of TRP1 RI Circle requires the products of the same genes (CDC28, CDC4, CDC7, and CDC8) required for replication of chromosomaL DNA. Like chromosomal DNA, its replication does not occur in cells arrested in the B1 phase of the cell cycle by incubation with the yeast pheromone alpha-factor. In addition, TRP1 RI Circle DNA is organized into nucleosomes whose size and spacing are indistinguishable from that of bulk yeast chromatin. These results indicate that TRP1 RI Circle has the replicative and structural properties expected for an origin of replication from yeast chromosomal DNA. Thus, this plasmid is a suitable model for further studies of yeast DNA replication in both cells and cell-free extracts.

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Cloning of promoter-like sequences from Lactobacillus paracasei subsp. paracasei CG11 and their expression in Escherichia coli, Lactococcus lactis, and Lactobacillus reuteri

Canadian Journal of Microbiology ◽

10.1139/m94-165 ◽

1994 ◽

Vol 40 (12) ◽

pp. 1043-1050 ◽

Cited By ~ 17

Author(s):

Gordana Djordjevic ◽

Bojana Bojovic ◽

Ana Banina ◽

Ljubisa Topisirovic

Keyword(s):

Escherichia Coli ◽

Lactococcus Lactis ◽

Dna Sequences ◽

Lactobacillus Reuteri ◽

Lactobacillus Paracasei ◽

Translational Efficiency ◽

Broad Host Range ◽

Chromosomal Dna ◽

Base Pairs ◽

E Coli

Fragments of chromosomal DNA from Lactobacillus paracasei subsp. paracasei CG11 (formerly Lactobacillus casei CG11) capable of functioning as promoters were isolated using the broad host range, promoter-probe vector pGKV210. Five such fragments designated P61, P79, P80, P116, and P144 were completely sequenced and analyzed. Fragment P61 had the highest transcriptional efficiency in Escherichia coli and Lactobacillus reuteri whereas P80 was the most active in Lactococcus lactis. In general, the orders of the transcriptional strengths were almost identical in E. coli and Lactobacillus reuteri but different from that in Lactococcus lactis. Mapping of the 5′ end of cat mRNA showed that different regions of fragments P79 and P144 were used as promoters in Lactococcus lactis than in E. coli and Lactobacillus reuteri. Analysis of these DNA sequences revealed that the putative −35 and −10 hexanucleotides resembled those of E. coli, Bacillus subtilis, and lactococci. The spacing between these two hexanucleotides and between the putative −10 hexanucleotide and the transcriptional start point (A residues predominated) ranged from 17 to 18 base pairs and from 5 to 7 base pairs, respectively. Each of the cloned Lactobacillus paracasei CG11 promoter-like fragments contained an AT-rich sequence upstream of the putative −35 region (from 60 to 73%).Key words: Lactobacillus, promoter-like sequences, transcriptional efficiency, translational efficiency.

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The refinement of species affiliation of some click beetles (Coleoptera: Elateridae) based on the molecular genetic analysis

Bulletin of the Karaganda University. “Biology, medicine, geography Series” ◽

10.31489/2021bmg1/14-19 ◽

2021 ◽

Vol 101 (1) ◽

pp. 14-19

Author(s):

T.M. Bragina ◽

◽

D.T. Konysbayeva ◽

M.M. Rulyeva ◽

M.A. Bobrenko ◽

...

Keyword(s):

Genetic Analysis ◽

Dna Sequences ◽

Sandy Loam ◽

Molecular Genetic ◽

Molecular Genetic Analysis ◽

Nucleotide Sequences ◽

Time Data ◽

Click Beetles ◽

Gene Coi ◽

University Of Oslo

The article is devoted to data on the approbation of modern methods and the refinement of species affiliation of soil-inhabiting larvae of click beetles (wireworm) based on the molecular genetic analysis (DNAbarcoding). For the first time, data obtained on the complete identity of certain DNA sequences of a number of species of click beetles living in the Kostanay Region (Kazakhstan) — dangerous pests of agricultural crops. At the same time, the World genetic bank (GenBank) did not find identical DNA sequences of decoded DNA nucleotide sequences for a number of studied specimens. The basis for this study was the materials collected in 2018 in the subzone of ordinary black earth on sandy loam soils (Mendykarinsky district). The selection of larvae was carried out by the method of standard soil-zoological samples. The fixation and storage of the selected click beetles larvae was carried out according to the method of preparing samples for molecular genetic analysis with fixation in 96 % alcohol. After the classic identification of the taxonomic position of the collected specimens, the species were identified by genetic analysis on the nucleotide sequence of the cytochrome C oxidase I subunit gene (COI). The assembly and decoding of the DNA nucleotide sequences of the studied samples were carried out using the programs «Codon Code Aligner» and «MEGA-X». As a result of the work carried out in the DNA laboratory of the Museum of Natural History (University of Oslo, Norway), it was possible to identify the complete identity of the DNA sequences of several mass species of click beetles, while a number of decoded DNA sequences of model specimens were absent in the genetic bank, which requires replenishment in it with new data.

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Molecular cytogenetic analysis and genetic counseling: a case report of eight 46,XX males and a literature review

Molecular Cytogenetics ◽

10.1186/s13039-019-0456-y ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 1

Author(s):

Fagui Yue ◽

Hongguo Zhang ◽

Qi Xi ◽

Yuting Jiang ◽

Leilei Li ◽

...

Keyword(s):

Genetic Counseling ◽

Molecular Genetic ◽

Sex Reversal ◽

Normal Male ◽

Case Presentation ◽

Molecular Cytogenetic ◽

Combined Application ◽

Genetic Techniques ◽

Male Sex ◽

Xx Males

Abstract Background 46,XX male syndrome is a rare disorder that usually causes infertility. This study was established to identify the genetic causes of this condition in a series of 46,XX males through the combined application of cytogenetic and molecular genetic techniques. Case presentation We identified eight azoospermic 46,XX males who underwent infertility-related consultations at our center. They all presented normal male phenotypes. In seven of the eight 46,XX males (87.5%), translocation of the SRY gene to the terminal short arm of the X chromosome was clearly involved in their condition, which illustrated that this translocation is the main mechanism of 46,XX sex reversal, in line with previous reports. However, one patient presented a homozygous DAX1 mutation (c.498G > A, p.R166R), which was not previously reported in SRY-negative XX males. Conclusions We proposed that this synonymous DAX1 mutation in case 8 might not be associated with the activation of the male sex-determining pathway, and the male phenotype in this case might be regulated by some unidentified genetic or environmental factors. Hence, the detection of genetic variations associated with sex reversal in critical sex-determining genes should be recommended for SRY-negative XX males. Only after comprehensive cytogenetic and molecular genetic analyses can genetic counseling be offered to 46,XX males.

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Organization of transcribed regions of chromatin

Philosophical Transactions of the Royal Society of London Series B Biological Sciences ◽

10.1098/rstb.1978.0036 ◽

1978 ◽

Vol 283 (997) ◽

pp. 343-357 ◽

Cited By ~ 14

Keyword(s):

Dna Sequences ◽

Messenger Rna ◽

Soluble Fraction ◽

Differential Sensitivity ◽

Chromosomal Dna ◽

Base Pairs ◽

Histone Protein ◽

Dnase Ii ◽

Specific Subset ◽

Active Subunits

The endonuclease DNase II preferentially attacks a limited and tissue-specific portion of chromosomal DNA. This material may be separated from the bulk of chromatin DNA by virtue of its solubility in 2 mM MgCl 2 . The Mg 2+ soluble fraction forms a specific subset of DNA sequences and is enriched four to sevenfold in sequences coding for cytoplasmic poly (A)-containing RNA and globin messenger RNA (in globin-producing cells). The bulk (70-90 %) of rapidly labelled RNA is found associated with the Mg 2 +-soluble fraction. Transcriptionally active, Mg 2+ -soluble chromatin is organized into repeating subunits of DNA (200 ± 5 base pairs) and histone. Mg 2 +-soluble active subunits differ from the subunits or nucleosomes of non-transcribed regions in many respects: namely, chemical composition (non-histone protein and RNA), sedimentation properties, differential sensitivity to DNase I and the single-strand-specific nuclease S1, and optical melting behaviour. These results suggest that chromatin subunits adopt a new configuration during the process of transcription.

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