scholarly journals Tolerance to Citrus mosaic virus in Transgenic Trifoliate Orange Lines Harboring Capsid Polyprotein Gene

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 865-868 ◽  
Author(s):  
Toru Iwanami ◽  
Tokurou Shimizu ◽  
Takao Ito ◽  
Toshio Hirabayashi
Keyword(s):  
Mosaic Virus ◽  
Binary Vector ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Poncirus Trifoliata ◽  
Trifoliate Orange ◽  
Chain Reaction ◽  
Southern Blot Hybridization Analysis ◽  
Transgenic Lines ◽  
Polymerase Chain

Trifoliate orange plants (Poncirus trifoliata) were transformed with a binary vector containing the capsid polyprotein (pCP) gene of Citrus mosaic virus (CiMV) via Agrobacterium tumefaciens LBA4404. Transformation was performed on the epicotyl segments obtained from a young seedling that was grown in the dark. Southern blot hybridization analysis showed that the transgene was stable in the transgenic lines after regeneration and propagation by grafting. Transgenic lines were screened for tolerance to CiMV by mechanical inoculation. Infection was monitored 30, 60, 90, and 120 days after inoculation by reverse transcription-polymerase chain reaction. The transgenic line 24 had the lowest infection rate (7.1%) at 60 days after inoculation, in contrast to that of nontransgenic plants (65.1%).The response of other lines to inoculation ranged from susceptibility to moderate tolerance.

scholarly journals Absence of the t(2;5) in Hodgkin's disease [see comments]

Blood ◽  
1995 ◽  
Vol 85 (10) ◽  
pp. 2845-2847 ◽  
Author(s):  
LM Weiss ◽  
JR Lopategui ◽  
LH Sun ◽  
OW Kamel ◽  
CH Koo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Hodgkin's Disease ◽  
Hodgkin’S Disease ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Large Cell ◽  
Common Finding ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Hodgkin's Lymphomas

The cytogenetics of Hodgkin's disease (HD) is poorly understood. However, a t(2;5) is a common finding in CD30+ anaplastic large cell lymphoma (ALCL), a neoplasm thought by some to be closely related to HD. Recently, the t(2;5) has been cloned and found to represent fusion of the NPM gene with the ALK gene. Using Southern blot hybridization, one group has reported finding rearrangements of NPM in a proportion of cases of both ALCL and HD. In the current study, we used a highly sensitive reverse transcriptase-polymerase chain reaction methodology to analyze 34 cases of HD for the t(2;5). We were unable to find polymerase chain reaction evidence for the t(2;5) in any of the cases of HD, a result significantly different from our previous study of CD30+ non-Hodgkin's lymphomas (P < .02) including ALCL (P < .04), using identical methods. Our results do not support the hypothesis that the t(2;5) represents a common chromosomal abnormality for both HD and ALCL.

Pitfalls in the epidemiologic classification of human papillomavirus types associated with cervical cancer using polymerase chain reaction: driver and passenger

2008 ◽  
Vol 18 (5) ◽  
pp. 1042-1050 ◽  
Author(s):  
T. Matsukura ◽  
M. Sugase
Keyword(s):  
Cervical Cancer ◽  
Polymerase Chain Reaction ◽  
Human Papillomavirus ◽  
Large Scale ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Cervical Lesions ◽  
Chain Reaction ◽  
Polymerase Chain

Cervical cancer is a common malignancy in women worldwide, and it has now been established that the human papillomavirus (HPV) is both necessary and causal for these lesions. HPV itself is both ubiquitous and markedly heterogeneous but can nevertheless be classified as either a high-risk type or a low-risk type based upon its frequency of detection in cervical cancer. Given that the association between HPV and cervical cancer is causal, the classification of this virus has been strengthened by large-scale epidemiologic studies and is widely accepted across many disciplines. It is evident, however, that cervical cancer is frequently associated with multiple HPV types. Therefore, it is crucial to distinguish causal types of HPV (drivers) from noncausal types (passengers) in cervical lesions. In this review, we highlight the current pitfalls of using polymerase chain reaction methods instead of Southern blot hybridization for detecting HPV and discuss the distinction between driver and passenger HPVs with regard to the viral type, the length of the viral genome, and the levels of viral DNA associated with cervical cancer. Finally, we newly propose three categories of HPV instead of two risk groups, based on similarities between viral genes

scholarly journals Multiple Citrus Viroids in Citrus from Japan and Their Ability to Produce Exocortis-Like Symptoms in Citron

2002 ◽  
Vol 92 (5) ◽  
pp. 542-547 ◽  
Author(s):  
Takao Ito ◽  
Hiroyuki Ieki ◽  
Katsumi Ozaki ◽  
Toru Iwanami ◽  
Kenji Nakahara ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Poncirus Trifoliata ◽  
Trifoliate Orange ◽  
Chain Reaction ◽  
Hop Stunt Viroid ◽  
Citrus Viroids ◽  
Citrus Medica ◽  
Polymerase Chain ◽  
Scaling Characteristic ◽  
Citrus Trees

Sequential polyacrylamide gel electrophoresis analyses showed many viroid-like RNAs in samples collected from citrus trees in Japan. Reverse transcription polymerase chain reaction and sequencing analyses of the amplified fragments verified that they were derived from variants of six citrus viroids, Citrus exocortis viroid (CEVd), Citrus bent leaf viroid (CBLVd) including CVd-I-LSS (a distinct variant of CBLVd), Hop stunt viroid, Citrus viroid III, Citrus viroid IV, and Citrus viroid OS. The samples induced symptoms with variable severity in Arizona 861-S1 ‘Etrog’ citrons (Citrus medica L.) likely due to the varying accumulation patterns produced by the different viroids. Some of the symptoms caused by the samples harboring the citrus viroids other than CEVd were as severe as those caused by CEVd. Some source citrus trees showing the severe bark scaling characteristic of exocortis disease in trifoliate orange (Poncirus trifoliata (L.) Raf.) rootstocks contained only citrus viroids other than CEVd in complex. This indicates that certain exocortis-like diseases in Japan were caused by some combination of citrus viroids not including CEVd.

Detection of Theileria annulata in cattle and vector ticks by PCR using the Tams1 gene sequences

Parasitology ◽  
2000 ◽  
Vol 120 (3) ◽  
pp. 245-254 ◽  
Author(s):  
E. KIRVAR ◽  
T. ILHAN ◽  
F. KATZER ◽  
P. HOOSHMAND-RAD ◽  
E. ZWEYGARTH ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Theileria Annulata ◽  
Babesia Bigemina ◽  
Chain Reaction ◽  
Hyalomma Anatolicum Anatolicum ◽  
Merozoite Surface Antigen ◽  
Base Pair Fragment ◽  
Polymerase Chain ◽  
Pair Fragment

A Polymerase Chain Reaction (PCR) and Southern blot hybridization for the detection of Theileria annulata are described. The PCR used primers amplifying a 785 base-pair fragment of the T. annulata gene which encodes the 30 kDa major merozoite surface antigen, Tams1. The sensitivity of the PCR in bovine blood was 1 piroplasm in 1 μl of blood. T. buffeli, T. parva, Babesia bigemina, B. bovis and B. divergens were not detected. The PCR detected down to 1 infected acinus/tick in resting and partially fed adult Hyalomma anatolicum anatolicum ticks and was negative for T. lestoquardi and T. equi, which are transmitted by this tick but are not infective to cattle. The specificity of the PCR was checked using 30 stocks of T. annulata, all of which were detected. Three stocks of T. lestoquardi, 4 of T. equi and 1 each of T. buffeli, T. parva, B. bigemina, B. bovis and B. divergens were used to ascertain there were no cross-reactions. A nested PCR using separate primers for the first reaction and the same primers for the second reaction detected T. annulata to the same sensitivity and specificity in saponin-extracted DNA samples stored for long periods at −20 °C.

scholarly journals Characterization of plasmid-mediated quinolone resistance by the qnrS gene in Escherichia coli isolated from healthy chickens and pigs

2009 ◽  
Vol 54 (No. 10) ◽  
pp. 473-482 ◽  
Author(s):  
H.-C. Kuo ◽  
C.-C. Chou ◽  
C. Tu ◽  
S.-R. Gong ◽  
C.-L. Han ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Quinolone Resistance ◽  
E Coli ◽  
Sequence Variations ◽  
Southern Blot Hybridization Analysis ◽  
Ciprofloxacin Resistance ◽  
Polymerase Chain

The prevalence of <I>qnr</I> and <I>qepA</I> genes in 660 <I>Escherichia coli</I> isolates was investigated in healthy animals from 30 pig farms and 30 chicken farms in Taiwan from January 2005 to February 2006 by the polymerase chain reaction. The <I>qnrS</I> gene, but not <I>qnrA, qnrB, </I> and <I>qepA</I> were detected in 12/360 pig isolates (3.33%) and in 6/300 chicken isolates (2%). Southern blot hybridization analysis indicated that <I>qnrS</I> was located on plasmids ranging in size from 50–165 kb. Eleven of the 18 <I>qnrS</I> positive isolates which showed a high ciprofloxacin resistance phenotype (minimum inhibitory concentration ≥ 8 mg/l) also had amino acid sequence variations in chromosomal quinolone resistance-determining regions of <I>gyrA</I> and <I>parC</I>. Only two <I>qnrS</I>-positive isolates carried the <I>aac(6’)-Ib-cr</I>variant that mediates FQ acetylation. For the high percentage resistance of cephalosporins, the<I> bla</I><sub>CTX-M</sub> gene was also examined in <I>qnrS</I>-positive isolates. The <I>bla</I><sub>CTX-M</sub> gene was detected in fifteen isolates (15/18, 83.3%) of which 12 isolates were <I>bla</I><sub>CTX-M-1</sub> and three isolates were <I>bl</I><sub>CTX-M-15</sub>. This study demonstrated a close linkage between the <I>qnrS</I> gene and <I>bla</I><sub>CTX-M-1</sub>, suggesting CTX-M and Qnr-based mechanisms might be co-emerging in <I>E. coli</I> strains isolated from healthy chickens and pigs under selective pressure of quinolone and cephalosporine administration.

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