scholarly journals Simultaneous and independent detection of C9ORF72 alleles with low and high number of GGGGCC repeats using an optimised protocol of Southern blot hybridisation

2013 ◽  
Vol 8 (1) ◽  
pp. 12 ◽  
Author(s):  
Vladimir L Buchman ◽  
Johnathan Cooper-Knock ◽  
Natalie Connor-Robson ◽  
Adrian Higginbottom ◽  
Janine Kirby ◽  
...  
Keyword(s):  
Southern Blot ◽  
Blot Hybridisation ◽  
Southern Blot Hybridisation

Evidence of HPV16 integration in low- and high-grade cervical lesions that regress demonstrated by multiple displacement amplification and Southern blot hybridisation

2007 ◽  
Vol 61 (4) ◽  
pp. 541-543 ◽  
Author(s):  
M F Evans ◽  
C Stewart-Crawford Adamson ◽  
K Cooper
Keyword(s):  
Southern Blot ◽  
Multiple Displacement Amplification ◽  
Intraepithelial Neoplasia ◽  
Low Grade ◽  
Physical Status ◽  
Cervical Lesions ◽  
Cytological Data ◽  
Blot Hybridisation ◽  
Southern Blot Hybridisation

The prevalence and significance of human papillomavirus (HPV) integration among different grades of cervical lesions is uncertain. In this study, HPV physical status was examined by the combination of multiple displacement amplification (MDA) with Southern blot hybridisation (SBH). DNA extracts from 95 cervical cytology samples (NILM, ASC-US, LSIL, ASC-H, HSIL) were subject to whole genome amplification by MDA followed by SBH with [α-32P]-labelled HPV probes. Mixed HPV16 episomal/integrant sequences were detected in three ASC-US patients (two diagnosed with benign changes and one with cervical intraepithelial neoplasia (CIN)2/3 after biopsy follow-up), one ASC-H patient with CIN2/3 histological diagnosis, and one HSIL patient with benign changes. Additional follow-up cytological data available for three of these patients demonstrated series of lesion-free samples. The data support the view that integration can occur in low-grade lesions and that lesions with mixed episomal/integrant HPV can regress.

Southern blot and dot blot hybridisation compared to PCR for the detection of human papillomavirus DNA in biopsies of the uterine cervix from women with dysplasia

1993 ◽  
Vol 1 (3) ◽  
pp. 187-194 ◽  
Author(s):  
Shujun Chen ◽  
Christopher K. Fairley ◽  
Sepehr N. Tabrizi ◽  
Michael A. Quinn ◽  
Suzanne M. Garland
Keyword(s):  
Human Papillomavirus ◽  
Southern Blot ◽  
Uterine Cervix ◽  
Dot Blot ◽  
Blot Hybridisation ◽  
Papillomavirus Dna ◽  
Dot Blot Hybridisation

Sequence and expression of a stem-abundant caffeic acid O-methyltransferase cDNA from perennial ryegrass (Lolium perenne)

1998 ◽  
Vol 25 (2) ◽  
pp. 225 ◽  
Author(s):  
Fiona M. McAlister ◽  
Colin L. D. Jenkins ◽  
John M. Watson
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Caffeic Acid ◽  
Lolium Perenne ◽  
Perennial Ryegrass ◽  
Leaf Sheath ◽  
Predicted Amino Acid Sequence ◽  
Degenerate Oligonucleotide ◽  
Blot Hybridisation ◽  
Southern Blot Hybridisation

A perennial ryegrass (Lolium perenne) cDNA library was screened with a PCR-amplified comt DNA fragment generated from ryegrass cDNA template using degenerate oligonucleotide primers. A full-length cDNA (LpeComt1) was isolated and shown to encode a caffeic acid O-methyltransferase (COMT) enzyme by analysis of its predicted amino acid sequence. This cDNA was expressed in E.coli and COMT activity was demonstrated in bacterial extracts. The predicted amino acid sequence of the perennial ryegrass COMT was highly similar (88%) with that of a COMT isolated from maize. Southern blot hybridisation analysis using this cDNA indicated that it is a member of a small gene family comprising at least two genes in perennial ryegrass. The LpeComt1 gene is strongly expressed in stem tissue, but not obviously in shoot, leaf blade, leaf sheath, flower or root tissue.

Survival of bacteria on wood and plastic particles: Dependence on wood species and environmental conditions

Holzforschung ◽  
2005 ◽  
Vol 59 (1) ◽  
pp. 72-81 ◽  
Author(s):  
Annett Milling ◽  
Rolf Kehr ◽  
Alfred Wulf ◽  
Kornelia Smalla
Keyword(s):  
Wood Species ◽  
Antibacterial Effect ◽  
The Other ◽  
Marker Genes ◽  
Bacterial Dna ◽  
E Coli ◽  
Hygroscopic Properties ◽  
Independent Analysis ◽  
Blot Hybridisation ◽  
Southern Blot Hybridisation

AbstractThe survival of two hygienically relevant bacteria,Escherichia colipIE639 andEnterococcus faecium, was followed on wooden sawdust of seven different European woods (pine, spruce, larch, beech, maple, poplar, and oak) versus polyethylene chips by using cultivation-dependent and molecular-based methods in parallel. The survival of the bacteria on wood was dependent on various factors such as the wood species, the type of the inoculated bacterium, the ambient temperature, and humidity. The bacterial titre decreased fastest on pine followed by oak compared to the other woods and plastic. Cultivation-independent analysis employing DNA extraction, Southern blot hybridisation, and PCR-based detection of marker genes of the test bacteria confirmed this result. The decline in bacterial numbers correlated with the decrease of bacterial DNA in the samples. Amounts of DNA ofE. coliandE. faeciumrecovered from pine and oak-wood sawdust were generally lower compared to the other woods and plastic.The presented study shows that pine and oak exhibit substantially better hygienic performance than plastic and indicates an antibacterial effect caused by a combination of the hygroscopic properties of wood and the effect of wood extractives.

Genetic transformation in commercial Tasmanian cultivars of opium poppy, Papaver somniferum, and movement of transgenicpollen in the field

2003 ◽  
Vol 30 (10) ◽  
pp. 1045 ◽  
Author(s):  
Julie A. Chitty ◽  
Robert S. Allen ◽  
Anthony J. Fist ◽  
Philip J. Larkin
Keyword(s):  
Transgenic Plants ◽  
Somatic Embryos ◽  
Papaver Somniferum ◽  
Opium Poppy ◽  
Weed Species ◽  
Mendelian Segregation ◽  
Pat Gene ◽  
Blot Hybridisation ◽  
Bottom Cooling ◽  
Southern Blot Hybridisation

We report a new transformation protocol for the pharmaceutically important opium poppy, Papaver somniferum L.; the protocol allows transformation for the first time of high yielding commercial cultivars. The method involves Agrobacterium tumefaciens infection of hypocotyl explants, followed by the production of antibiotic- or herbicide-resistant embryogenic callus and the subsequent induction of somatic embryos and plants. Key elements of the improvement are the use of buffering agents to stabilise medium pH and bottom-cooling of the cultures. Transformation was verified by PCR and Southern blot hybridisation. Transcription of transgenes was confirmed by RT–PCR and product sequencing. Expression of transgenes was detected by histochemical GUS staining, phosphinothricin acetyltransferase (PAT) enzyme assays for bar and pat genes, and western analysis of transgenic sunflower seed albumin protein. Expression of various transgenes was detected in stem, leaf, seed, capsule and latex. The pat gene was demonstrated to be stably inherited to the T2 generation and to confer phosphinothricin (PPT) herbicide resistance. Most T0 plants showed normal morphology, were self-fertile and the transgenes displayed the expected Mendelian segregation. The percentage of explants producing somatic embryos that developed into plantlets able to be transplanted to soil, ranged from 6–11% in two Tasmanian cultivars.A field trial using pat transgenic plants was designed to estimate the frequency of hybridisation at various distances into buffer rows of non-transgenic poppies and to related weed species, P. somniferum spp. setigerum and P. dubium. The frequency of hybridisation to completely compatible poppy fell sharply with distance, being 2.6% at 20 cm, 2.13% at 0.5 m and falling to 0% at 2.5 and 5 m. No hybridisation could be detected to two weed species under open pollination conditions, including the compatible P. somniferum spp. setigerum, when grown as close as 20 cm, despite flowering at the same time as the transgenic plants in the presence of foraging bees.

Detection and differentiation of phytoplasmas in Australia

1997 ◽  
Vol 48 (5) ◽  
pp. 535 ◽  
Author(s):  
R. I. Davis ◽  
B. Schneider ◽  
K. S. Gibb
Keyword(s):  
Sweet Potato ◽  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Aster Yellows ◽  
Site Analysis ◽  
Blot Hybridisation ◽  
Total Dna ◽  
Polymerase Chain ◽  
Southern Blot Hybridisation ◽  
Polymorphism Pattern

In a polymerase chain reaction (PCR) diagnostic test, phytoplasma (formerly known as plant-pathogenic mycoplasma-like organism or MLO) ribosomal DNA was detected in total DNA extracts prepared from 56 out of 63 plants collected from geographically diverse locations across Australia. The list of phytoplasma hosts consisted of 38 different species in 16 different families. Restriction site analysis of the PCR-amplified DNA accessions was used to divide the phytoplasmas into 2 groups. The majority of the tomato big bud group and sweet potato little leaf group phytoplasmas were closely related to a phytoplasma originally obtained from Crotalaria in Thailand, which is a member of the faba bean phyllody strain cluster. In contrast, phytoplasmas associated with Australian grapevine yellows and papaya dieback were most similar to members of the aster yellows strain cluster. Twelve phytoplasmas were compared by Southern blot hybridisation with DNA cloned from the sweet potato little leaf phytoplasma strain V4. The restriction fragment length polymorphism pattern of all phytoplasmas compared was identical except for 2 sweet potato little leaf phytoplasmas.

Expression of c-fos and c-jun proto-oncogenes by ovine preimplantation embryos

Zygote ◽  
1996 ◽  
Vol 4 (3) ◽  
pp. 211-217 ◽  
Author(s):  
Bin Wu
Keyword(s):  
Regulation Of Gene Expression ◽  
Preimplantation Embryos ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Mrna Transcripts ◽  
Pcr Products ◽  
Blot Hybridisation ◽  
Hybridisation Analysis ◽  
Polymerase Chain ◽  
Southern Blot Hybridisation

SummaryThe c-fos and c-jun proto-oncogenes are involved in the regulation of gene expression, cell proliferation, differentiation and tumorigenesis. Embryogenesis, like tumorigenesis, involves dramatic cell growth, cleavage and differentiation processes. Activation of the c-fos and c-jun proto-oncogenes in sheep conceptuses during the period of rapid growth and elongation was examined using reverse transcription and polymerase chain reaction (RT-PCR). The specificity of PCR products was determined by Southern blot hybridisation analysis with a non-radioactive DNA probe. A band corresponding to a 507 bp fragment (predicted amplified c-fos gene cDNA product) was observed in 3 of 9 day-13, 1 of 4 day-14 and 1 of 2 day-16 embryos. Meanwhile, a 400 bp c-jun transcript was aslo detected in 1 or 2 day-12, 3 of 9 day-13 and 2 of 2 day-16 embroyos. These results suggest that mRNA transcripts of c-fos and c-jun proto-oncogenes were specifically expressed during the period of ovine embryonic elongation and may have a possible role in preimplantation embryonic development of sheep.

Humán papillomavírus (HPV-16) kimutatása a colon laphámsejtes carcinomájából és annak nyirokcsomó-metastasisaibólr

2008 ◽  
Vol 61 (4) ◽  
pp. 225-229
Author(s):  
Gábor Bognár ◽  
Gábor István ◽  
György Ledniczky ◽  
Pál Ondrejka
Keyword(s):  
Human Papillomavirus ◽  
Southern Blot ◽  
Hpv 16

Absztrakt A humán papillomavírus (HPV) etiológiai szerepe számos laphámeredetű malignomában ismert. A colon laphámsejtes rákja (SCC) ritka betegség, etiológiája bizonytalan. Egy általunk operált SCC-eset kapcsán célunk volt kimutatni egy esetleges HPV-fertőzést. Egy 94 éves nőbeteget operáltunk passage-zavart okozó colon descendens tumor miatt. A szövettani vizsgálat SCC-t igazolt. Korábban kidolgozott módszerrel PCR- és Southern blot hybridisatiós technikával vizsgáltuk meg a daganatszövetet és az eltávolított nyirokcsomókat. Az eset kapcsán átnéztük az SCC-re vonatkozó irodalmat. A malignomákban leggyakrabban előforduló HPV-típusok (16, 18) közül a HPV-16 jelenlétét tudtuk igazolni a SCC-ben és a környező 9 nyirokcsomóból 4-ben. Ezek közül 2 metastaticus volt. Egy SCC-s betegnél sikerült HPV-16-fertőzöttséget kimutatni, a primer tumorból és a metastaticus nyirokcsomók mellett további környező nyirokcsomókból. Ilyen tanulmányt tudomásunk szerint még senki nem közölt.

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