Characterisation of isolates and strains of citrus tristeza closterovirus using restriction analysis of the coat protein gene amplified by the polymerase chain reaction

1993 ◽  
Vol 44 (2-3) ◽  
pp. 305-317 ◽  
Author(s):  
Michael Gillings ◽  
Patricia Broadbent ◽  
James Indsto ◽  
Richard Lee
Keyword(s):  
Polymerase Chain Reaction ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Restriction Analysis ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Citrus Tristeza Closterovirus ◽  
Citrus Tristeza

scholarly journals Reverse-Transcription Polymerase Chain Reaction Detection of Citrus Tristeza Virus in Aphids

Plant Disease ◽  
1997 ◽  
Vol 81 (9) ◽  
pp. 1066-1069 ◽  
Author(s):  
Prem Mehta ◽  
R. H. Brlansky ◽  
S. Gowda ◽  
R. K. Yokomi
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Citrus Tristeza Virus ◽  
Protein Gene ◽  
Aphid Species ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tristeza Virus

A rapid and simple reverse-transcription polymerase chain reaction (RT-PCR) method was developed for the detection of citrus tristeza virus (CTV) in three aphid species. Seven CTV isolates from a worldwide isolate collection were used for aphid acquisition feeding by three aphid species. These included the most efficient CTV vector, the brown citrus aphid, Toxoptera citricida; the melon aphid, Aphis gossypii; and the green peach aphid, Myzus persicae, a non-vector for CTV. A short procedure for nucleic acid extraction from single or groups of aphids was developed. Nucleic acid extracts from 1, 3, 5, and 10 aphids with acquisition-access periods of 24 and 48 h were reverse transcribed and amplified using primers for the coat protein gene of the Florida B3 (T-36) isolate of CTV. PCR-amplified fragments of approximately 670 bp were obtained from all the isolates tested and the amplified product from the aphids fed on citrus infected with isolate B3 was confirmed as the CTV coat protein gene by digesting with various restriction enzymes. This technique will be useful in investigations of CTV-vector-plant interactions and CTV epidemiology.

scholarly journals PENGGUNAAN PELACAK NONRADIOAKTIF (Digoxigenin-DNA Probe) UNTUK MENDETEKSI PEANUT STRIPE VIRUS

2001 ◽  
Vol 1 (2) ◽  
pp. 75-79
Author(s):  
Hasriadi Mat Akin
Keyword(s):  
Polymerase Chain Reaction ◽  
Coat Protein ◽  
Untranslated Region ◽  
Protein Gene ◽  
Dna Probe ◽  
Nuclear Inclusion ◽  
Chain Reaction ◽  
Peanut Stripe Virus ◽  
Polymerase Chain ◽  
Labeled Probe

The use of nonradioactive probe (Digoxigenin-DNA)  for  detection of peanut stripe virus.  The objective of this experiment was to develop the nonradioactive-labeled probe to detect peanut stripe virus (PStV) in peanut leaves and seeds. Digoxigenin labeled cDNA (dig-DNA probe) was synthesized from recombinant plasmid (pHS1.23) using polymerase chain reaction (PCR).  The probe containing 1.195 bp (base pair) corresponding to 3' termini, included part of NIb (nuclear inclusion body) gene, coat protein gene, and 3' untranslated region of PStV genome was used to detect the existence of PStV in peanut leaves and seeds of infected peanut plants.  

Detection of the Glanzmann's Thrombasthenia Mutations in Arab and Iraqi-Jewish Patients by Polymerase Chain Reaction and Restriction Analysis of Blood or Urine Samples

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman
Keyword(s):  
Polymerase Chain Reaction ◽  
Base Pair ◽  
Restriction Analysis ◽  
Urine Samples ◽  
Chain Reaction ◽  
Base Pairs ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Base Pair Deletion ◽  
Polymerase Chain

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.

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