d(GAmiddle dotTC)n microsatellite DNA sequences enhance homologous DNA recombination in SV40 minichromosomes

Etoposide is one of the most widely used antineoplastics. Unfortunately, the same treatment schedules associated with impressive efficacy are associated with an increased risk of secondary acute myeloid leukemia (AML), which has prompted its withdrawal from some treatment regimens, thereby potentially compromising efficacy against the original tumor. Because etoposide-associated AML is characterized by site-specific illegitimate DNA recombination, we studied whether etoposide could directly cause site-specific deletions of exons 2 and 3 in the hprt gene. Human lymphoid CCRF-CEM cells were treated with etoposide for 4 hours, and DNA was isolated after subculturing. The deletion of exons 2 and 3 from hprt was assayed by a quantitative polymerase chain reaction (PCR) method. In the absence of etoposide treatment, the frequency of deletions of exons 2 and 3 was very low (5.05 x 10(-8)). After exposure to 10 mumol/ L etoposide, the frequency of the exon 2 + 3 deletion was increased immediately after and at 24 hours after etoposide treatment (65 to 89 x 10(-8)) and increased to higher levels (128 to 173 x 10(-8)) after 2 and 6 days of subculture (P < .001 overall). The frequency of the exon 2 + 3 deletion assessed at 6 days of subculture after 4 hours of 0, 0.25, 1, 2.5, 5, and 10 mumol/L etoposide treatment increased with etoposide concentration, ie, 5.05 x 10(-8), 89.2 x 10(-8), 108 x 10(-8), 142 x 10(-8), 163 x 10(-8), and 173 x 10(-8), respectively (P < .0001). Sequencing of a subset of amplified products confirmed the presence of DNA sequences at the breakpoints consistent with V(D)J recombination. By contrast, exon 2 + 3 deletions after etoposide treatment in the myeloid cell lines KG-1A and K562 showed no evidence of V(D)J recombinase in their genesis. We conclude that etoposide can induce the illegitimate site-specific action of V(D)J recombinase on an unnatural DNA substrate after a single treatment in human lymphoid cells.

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Genetic variation in the population of three Polish cattle breeds included into the programme of genetic resources protection and Holstein-Friesian breed, estimation on the basis of polymorphism of 24 microsatellite DNA sequences

AFRICAN JOURNAL OF BIOTECHNOLOGY ◽

10.5897/ajb12.1050 ◽

2012 ◽

Vol 11 (77) ◽

Cited By ~ 2

Author(s):

Wioletta Sawicka-Zugaj

Keyword(s):

Genetic Variation ◽

Genetic Resources ◽

Dna Sequences ◽

Microsatellite Dna ◽

Cattle Breeds ◽

Holstein Friesian

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The phylogeography and population genetics of Polyprion oxygeneios based on mitochondrial DNA sequences and microsatellite DNA markers

Fisheries Research ◽

10.1016/j.fishres.2015.08.009 ◽

2016 ◽

Vol 174 ◽

pp. 19-29 ◽

Cited By ~ 4

Author(s):

Henry S. Lane ◽

Jane E. Symonds ◽

Peter A. Ritchie

Keyword(s):

Population Genetics ◽

Mitochondrial Dna ◽

Dna Sequences ◽

Dna Markers ◽

Microsatellite Dna ◽

Microsatellite Dna Markers ◽

Polyprion Oxygeneios

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Repressing Integrase attachment site operation with CRISPR-Cas9 in E. coli

10.1101/110254 ◽

2017 ◽

Author(s):

Andrey Shur ◽

Richard M. Murray

Keyword(s):

Dna Sequences ◽

Dna Recombination ◽

Attachment Site ◽

Memory Systems ◽

Rna Sequences ◽

Guide Rna ◽

Future Goals ◽

Attachment Sites ◽

Synthetic Cell ◽

Specific Pair

AbstractSerine integrases are bacteriophage proteins responsible for integrating the phage genome into that of the host. Synthetic biologists have co-opted these proteins into useful tools for permanent DNA logic, utilizing their specific DNA recombination abilities to build synthetic cell differentiation and genetic memory systems. Each integrase has a specific pair of DNA sequences (attP/attB sites) that it recombines, but multiple identical sites can result in unpredictable recombination. We have developed a way to control integrase activity on identical attP/attB sites by using catalytically dead Cas9 (dCas9) as a programmable binding protein that can compete with integrase for binding to specific attachment sites. Utilizing a plasmid that contains two identical Bxb1 attP sites, integration can be repressed up to 8 fold at either one of the two attP sites when guide RNA and dCas9 are present. Guide RNA sequences that bind specifically to attB, or either of two attP sites, have been developed. Future goals are to utilize this technology to construct larger and more complex integrase logic circuits.

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Polymorphism of bovine microsatellite DNA sequences in the lowland European bison

ACTA THERIOLOGICA ◽

10.1007/bf03192589 ◽

2004 ◽

Vol 49 (4) ◽

pp. 449-456 ◽

Cited By ~ 7

Author(s):

Barbara Gralak ◽

Małgorzata Krasińska ◽

Cezary Niemczewski ◽

Zbigniew A. Krasiński ◽

Maciej Żurkowski

Keyword(s):

Dna Sequences ◽

Microsatellite Dna ◽

European Bison

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The Mycoplasma pneumoniae MPN490 and Mycoplasma genitalium MG339 Genes Encode RecA Homologs That Promote Homologous DNA Strand Exchange

Infection and Immunity ◽

10.1128/iai.00747-09 ◽

2009 ◽

Vol 77 (11) ◽

pp. 4905-4911 ◽

Cited By ~ 9

Author(s):

Marcel Sluijter ◽

Emiel B. M. Spuesens ◽

Nico G. Hartwig ◽

Annemarie M. C. van Rossum ◽

Cornelis Vink

Keyword(s):

Mycoplasma Pneumoniae ◽

Dna Sequences ◽

Sequence Variation ◽

Antigenic Variation ◽

Bacterial Genome ◽

Amino Acid Level ◽

Dna Recombination ◽

Mycoplasma Genitalium ◽

Repeat Elements ◽

Dna Substrates

ABSTRACT The P1, P40, and P90 proteins of Mycoplasma pneumoniae and the MgPa and P110 proteins of Mycoplasma genitalium are immunogenic adhesion proteins that display sequence variation. Consequently, these proteins are thought to play eminent roles in immune evasive strategies. For each of the five proteins, a similar underlying molecular mechanism for sequence variation was hypothesized, i.e., modification of the DNA sequences of their respective genes. This modification is thought to result from homologous recombination of parts of these genes with repeat elements (RepMp and MgPar elements in M. pneumoniae and M. genitalium, respectively) that are dispersed throughout the bacterial genome. Proteins that are potentially involved in homologous DNA recombination have been suggested to be implicated in recombination between these repeat elements and thereby in antigenic variation. To investigate this notion, we set out to study the function of the RecA homologs that are encoded by the M. pneumoniae MPN490 and M. genitalium MG339 genes. Both proteins, which are 79% identical on the amino acid level, were found to promote recombination between homologous DNA substrates in an ATP-dependent fashion. The recombinational activities of both proteins were Mg2+ and pH dependent and were strongly supported by the presence of single-stranded DNA binding protein, either from M. pneumoniae or from Escherichia coli. We conclude that the MPN490- and MG339-encoded proteins are RecA homologs that have the capacity to recombine homologous DNA substrates. Thus, they may play a central role in recombination between repetitive elements in both M. pneumoniae and M. genitalium.

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Interspecies and intraspecies genetic diversity of Ongole Grade cattle and Madura cattle based on microsatellite DNA markers

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d190453 ◽

2018 ◽

Vol 19 (4) ◽

pp. 1593-1600

Author(s):

SUTARNO . ◽

NINA KURNIANINGRUM ◽

ELISA HERAWATI ◽

AHMAD DWI SETYAWAN

Keyword(s):

Genetic Diversity ◽

Dna Sequences ◽

Dna Markers ◽

Microsatellite Loci ◽

Microsatellite Dna ◽

Polymorphic Information Content ◽

Entire Genome ◽

Genetic Characteristics ◽

Microsatellite Dna Markers ◽

The Mean

Sutarno, Kurnianingrum N, Herawati E, Setyawan AD. 2018. Interspecies and intraspecies genetic diversity of Ongole Gradecattle and Madura cattle based on microsatellite DNA markers. Biodiversitas 19: 1593-1600. DNA microsatellite has been extensivelyemployed for estimating the degree of kinship between genotypes and improving the quality of cattle products. Microsatellite markersare short-patterned DNA sequences and repeated tandem (sequentially) with 2-5 nucleotide units scattering the entire genome. Thepurpose of this study was to investigate the genetic characteristics of inter and intraspecies of Ongole Grade cattle and Madura cattleusing microsatellite DNA markers. Blood samples from 20 individuals of each species were extracted by the method referring to WizardGenomic DNA Purification Kit (Promega, USA) and PCR amplification was performed using 5 microsatellite loci, i.e., BM1824,ETH225, INRA005, MM12, and TGLA227. Results of the genetic characteristics of both species were calculated using the POPGENEprogram version 1.31. The data suggest that there is a genetic diversity of inter and intraspecies of Ongole Grade cattle and Maduracattle. The average value of Shannon's Information Index (I) at all microsatellite loci for Ongole Grade cattle was 0.76 and for Maduracattle was 1.12. Meanwhile, the average interspecies I value was 1.03. The mean intraspecies Polymorphic Information Content (PIC) ofOngole Grade and Madura cattle was 0.43, and 0.63, respectively, and the mean interspecies PIC value was 0.57. The data altogethersuggest that all loci meet the standards as being informative markers in the assessment of genetic population because it has a PIC value>0.5 especially for intraspecies of Madura cattle.

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