scholarly journals Detection and molecular characterization of the Iris severe mosaic virus-Ir isolate from Iran

2015 ◽  
Vol 55 (3) ◽  
pp. 235-240 ◽  
Author(s):  
Masoud Nateqi ◽  
Mina Koohi Habibi ◽  
Akbar Dizadji ◽  
Shirin Parizad
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Mosaic Virus ◽  
Ornamental Plants ◽  
Rt Pcr ◽  
Chain Reaction ◽  
C Terminus ◽  
Coated Plate ◽  
Polymerase Chain

AbstractIris belongs to the Iridaceae family. It is one of the most important pharmaceutical and ornamental plants in the world. To assess the potyvirus incidence in natural resources of iris plants in Iran, Antigen Coated-Plate ELISA (ACP-ELISA) was performed on 490 symptomatic rhizomatous iris leaf samples, which detected the potyvirus in 36.7% of the samples. Genomic 3′ end of one mechanically non-transmitted potyvirus isolate, comprising a 3′ untranslated region (390 bp) and C-terminus of the coat protein (CP) gene (459 bp), was amplified by reverse transcription polymerase chain reaction (RT-PCR), which was ligated into pTG19-T vector. The nucleotide sequence of amplicons was compared with related sequences, using Blastn software available at NCBI GenBank, and showed the highest similarity withIris severe mosaic virus(ISMV) isolates. The nucleotide and deduced amino acid sequence of the CP C-terminus region was more than 83% identical with other ISMV isolates, therefore this isolate was designated as ISMV-Ir. This new ISMV isolate is closely related to the Chinese ISMV-PHz in phylogenetic analysis, based on the partial nucleotide and deduced amino acid sequence of the CP region. This is the first report of ISMV occurrence onIrisspp. in Iran.

scholarly journals First Detection and Characterization of Chrysanthemum virus B infecting Chrysanthemum in Thailand

2021 ◽  
Author(s):  
Salit Supakitthanakorn ◽  
Garnjana Wichitrakoonthavorn ◽  
Kaewalin Kunasakdakul ◽  
On-Uma Ruangwong
Keyword(s):  
Cultural Values ◽  
Ornamental Plants ◽  
Systemic Symptom ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Transmission Electron ◽  
Staining Methods ◽  
Polymerase Chain ◽  
Systemic Symptoms

Abstract Chrysanthemum is one of the important ornamental plants in worldwide due to its high economic and cultural values. Chrysanthemum leaves showed mosaic, ringspot, yellowing and mild mottle symptoms were observed and collected from cultivation areas in northern Thailand and used for detection of important viruses infecting chrysanthemum. Chrysanthemum virus B (CVB) was detected by reverse transcription polymerase chain reaction (RT-PCR) from samples showing yellowing and mild mottle symptoms. Sequences of the coat protein (CP) gene of two CVB isolates found in this study were sequenced and shared 93.15% homology with other CVB isolates from different countries deposited in GenBank. Biological indexing of these CVB found that they induced both local and systemic symptoms in tobacco plants while petunia displayed a systemic symptom. The particles of CVB were observed under transmission electron microscope (TEM), prepared by dip preparation and negative staining methods, showing slightly flexuous rod-shaped virions approximately 600–650 nm in length. To our knowledge, this is the first detection and study on molecular and biological characteristics of CVB infecting chrysanthemum in Thailand.

scholarly journals Nucleotide sequence of the coat protein gene of the Skierniewice isolate of plum pox virus (PPV).

1994 ◽  
Vol 41 (1) ◽  
pp. 87-95 ◽  
Author(s):  
K Wypijewski ◽  
T Malinowski ◽  
W Musiał ◽  
J Augustyniak
Keyword(s):  
Amino Acid ◽  
Nucleotide Sequence ◽  
Coat Protein ◽  
Amino Acid Sequence ◽  
Protein Gene ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Very High

The coat protein (CP) gene of the Skierniewice isolate of plum pox virus (PPV-S) has been amplified using the reverse transcription--polymerase chain reaction (RT-PCR), cloned and sequenced. The nucleotide sequence of the gene and the deduced amino-acid sequence of PPV-S CP were compared with those of other PPV strains. The nucleotide sequence showed very high homology to most of the published sequences. The motif: Asp-Ala-Gly (DAG), important for the aphid transmissibility, was present in the amino-acid sequence. Our isolate did not react in ELISA with monoclonal antibodies MAb06 supposed to be specific for PPV-D.

scholarly journals Human FLT3/FLK2 gene: cDNA cloning and expression in hematopoietic cells

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1110-1119 ◽  
Author(s):  
O Rosnet ◽  
C Schiff ◽  
MJ Pebusque ◽  
S Marchetto ◽  
C Tonnelle ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Tyrosine Kinase ◽  
B Cell ◽  
Cloning And Expression ◽  
Colony Stimulating Factor ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Stimulating Factor

The human FLT3 cDNA was cloned from a pre-B cell line and characterized. The deduced amino acid sequence shows that FLT3 codes for a receptor-type tyrosine kinase of 993 residues, presenting a strong similarity with the corresponding mouse FLT3/FLK2 protein as well as with the receptors for colony-stimulating factor 1 (CSF1R/FMS) and steel locus factor (SLFR/KIT). An analysis of the expression of the gene using amplification of reverse transcribed FLT3 mRNA by polymerase chain reaction shows that FLT3 is expressed in various lymphohematopoietic cells and tissues, including a series of immature cell lines and leukemias of lymphocytic origin.

scholarly journals Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa

2018 ◽  
Vol 14 (3) ◽  
pp. 75
Author(s):  
Listihani Listihani ◽  
Tri Asmira Damayanti ◽  
Sri Hendrastuti Hidayat ◽  
Suryo Wiyono
Keyword(s):  
Ringspot Virus ◽  
Mosaic Symptom ◽  
Papaya Ringspot Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Specific Primers ◽  
Central Java ◽  
Dna Fragment ◽  
Polymerase Chain

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody.  Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java.  Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA).  Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively.  Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing.  DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W.  Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively.  This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia.  This is the first report of PRSV-P infecting cucumber in Indonesia.

scholarly journals First Report of a Lettuce-Infecting Sequivirus in Chile

Plant Disease ◽  
2005 ◽  
Vol 89 (10) ◽  
pp. 1129-1129 ◽  
Author(s):  
R. Krause-Sakate ◽  
A. S. Jadão ◽  
A. C. Firmino ◽  
M. A. Pavan ◽  
F. M. Zerbini ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mosaic Virus ◽  
Plant Viruses ◽  
Yellow Mosaic Virus ◽  
Mixed Infections ◽  
Replicase Gene ◽  
Rt Pcr ◽  
Chain Reaction ◽  
First Report ◽  
Polymerase Chain

Sequiviruses are isometric aphidborne plant viruses. Dandelion yellow mosaic virus (DaYMV), genus Sequivirus, was isolated from dandelion and lettuce in Europe. Lettuce mottle virus (LeMoV), a putative sequivirus, is often found in mixed infections with Lettuce mosaic virus (LMV) in Brazil (3). DaYMV, LeMoV and LMV cause similar mosaics in field-grown lettuce. Differences in biology and sequence suggest that DaYMV and LeMoV are distinct species (2). Forty-two and 101 lettuce samples with mosaic symptoms collected from two locations near Santiago during a survey of lettuce viruses in Chile in 2002 and 2003, respectively, were analyzed for the presence of LeMoV using reverse transcription polymerase chain reaction (RT-PCR). Total RNA was extracted (1) and used for RT-PCR with the specific LeMoV primers pairs Lmo3 (5′ ACATGAGCACTAGTGAGG 3′) and Lmo4 (5′ AGATAGAGCCGTCT GGCG 3′) (2). One of the 42 and three of the 101 samples produced the expected 300-bp fragment. Isometric particles of 30 nm diameter, typical of a sequivirus, were visualized by transmission electron microscopy. These samples were tested using RT-PCR for the presence of LMV and Cucumber mosaic virus (CMV), but no mixed infections were observed. One isolate, Ch36, was reamplified with the degenerate primer pairs DALE 1 (5′ GARTTCAACATGCACGCCAG 3′) and DALE 2 (5′ TTTTTCTCCCCATYCGTCAT 3′) which amplify part of the putative replicase gene (2) and produced a 563-bp fragment that was cloned on pGEM-T Easy (Promega, Madison, WI) and sequenced. The Ch36 product (EMBL Accession No. AM039965) showed 97% amino acid identity with LeMoV from Brazil, 79% with DaYMV, 72% with the sequivirus Parsnip yellow fleck virus, and 34% with the waikavirus Maize chlorotic dwarf virus. To our knowledge, this is the first report of a sequivirus in field lettuce in Chile, and although the virus was found at low incidence, this report extends the range of LeMoV to the western side of the Cordillera de Los Andes. The impact of LeMoV needs to be further analyzed in Chile, Brazil, and possibly other South American countries. References: (1) Y. D. Bertheau et al. DNA amplification by polymerase chain reaction (PCR) 1998. In: Methods for the Detection and Quantification of Erwinia carotovora subsp. atroseptica on potatoes. M. C. N. Perombelon and J. M. van der Wolff, eds. Scott. Crop Res. Inst. Occasional Publ., Dundee, 1998. (2) A. S. Jadão. Caracterização parcial e desenvolvimento de oligonucleotídeos específicos para detecção de sequivirus infectando alface. Ph.D. thesis. FCA-UNESP-Botucatu, Brazil, 2004. (3) O. Stangarlin et al. Plant Dis. 84:490, 2000.

scholarly journals Sequence of the S10 cDNA from 'McIntosh' Apple and a PCR-digestion Identification Method

HortScience ◽  
2002 ◽  
Vol 37 (1) ◽  
pp. 187-190 ◽  
Author(s):  
Kentaro Kitahara ◽  
Shogo Matsumoto
Keyword(s):  
Polymerase Chain Reaction ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Restriction Enzyme ◽  
Chain Reaction ◽  
Specific Primers ◽  
Amino Acid Sequence Level ◽  
Identification Method ◽  
Polymerase Chain

An S-allele cDNA was cloned from pistils of 'McIntosh' apple (Malus ×domestica Borkh.). The allele, designated Si in Japan and S10 in Europe, is an S-RNase that is very similar (94%) to the S3-RNase at the deduced amino acid sequence level. This allele can be detected by amplification using the polymerase chain reaction (PCR) and specific primers, followed by digestion with restriction enzyme EheI. The S10 allele was discovered in 'Empire', 'Maypole', 'Shinano Red', 'Spencer', and 'Vista Bella'. The S-allele cDNAs sequenced to date are listed with their Japanese and European designations.

Cloning and sequencing of a cDNA encoding an ovine oestrus-associated oviducal protein

1996 ◽  
Vol 8 (2) ◽  
pp. 305 ◽  
Author(s):  
JT Marshall ◽  
CD Nancarrow ◽  
AG Brownlee
Keyword(s):  
Amino Acids ◽  
Polymerase Chain Reaction ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Northern Blot ◽  
Mature Protein ◽  
Chain Reaction ◽  
Cloning And Sequencing ◽  
Glycosylation Sites ◽  
Polymerase Chain

Ovine oestrus-associated oviducal glycoprotein (oEGP) is synthesized and secreted specifically by the ampullary region of the ovine oviduct during the peri-ovulatory stages of the oestrous cycle. A cDNA that encodes oEGP was isolated and sequenced. Isolation of oEGP was achieved using the polymerase chain reaction (PCR) with primers based on a bovine oestrus-associated oviducal glycoprotein cDNA (bOGP) sequence. A 1599-bp cDNA encodes, in part, a deduced 519-amino acid sequence of mature protein which carries two potential N-linked glycosylation sites. The deduced amino acid sequence is more than 95% identical to that of bOGP and more than 74% identical to the first 491 amino acids of human oestrogen-dependent oviducal glycoprotein (hOGP). Northern blot hybridizations of RNA from several sheep tissues detected mRNA (2.4 kb) only in an ampulla oviduct sample.

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