scholarly journals Evaluation of SDS-Polyacrylamide gel electrophoresis and immunostrip technique for detection of Tobacco mosaic virus and Cucumber mosaic virus in IRAQ

2011 ◽  
Vol 5 (2) ◽  
pp. 85-94
Author(s):  
Rakib A. Al-Ani ◽  
Mustafa A. Adhab
Keyword(s):  
Tobacco Mosaic Virus ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Positive Reaction ◽  
Total Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Band ◽  
Polyacrylamide Gel ◽  
Total Proteins ◽  
Winter Squash

his study was carried out to evaluate the efficiency of electrophoresis on SDS- poly acrylamide slap gel and immunostrip techniques for detection of Tobacco mosaic virus (TMV, genus Tobamovirus) and Cucumber mosaic virus (CMV, genus Cucumovirus, family Bromoviridae), compared with symptoms on diagnostic plants for the two viruses. The results obtained showed that the two methods were effective. The analysis of samples of purified CMV, total proteins from infected cucumber plants, and extracts from infected plants with or without chlorophyll, by electrophoresis on 10% polyacrylamide slap gel containing 0.1% SDS showed two bands of 24 and 26 kd in size, and absent in samples of total protein or extracts of healthy plants. These two proteins represent the coat protein (CP) of CMV. In addition, one 18 kd protein band appeared on SDS- polyacrylamide gel profile which represent the CP of TMV, when samples of purified virus, total protein of infected plants, and plant extracts with or without chlorophyll were analyzed. This band was absent in similar samples from healthy plants. The test of immunostrip specific for CMV showed positive reaction with extracts from melon, cucumber, winter squash, and zucchini infected plants. Similarly, a positive reaction with immunostrip specific for TMV appeared with extracts from tobacco, tomato infected with TMV. No reaction was obtained with healthy plants extract. These results were similar to those obtained from indicator plants for the two viruses.

Glycoproteins, enzymatic activities, and b proteins in intercellular fluid extracts from hypersensitive Nicotiana species infected with tobacco mosaic virus

1985 ◽  
Vol 63 (5) ◽  
pp. 928-931 ◽  
Author(s):  
Jean-Guy Parent ◽  
Richard Hogue ◽  
Alain Asselin
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Electrophoretic Pattern ◽  
Protein Band ◽  
Polyacrylamide Gel ◽  
Intercellular Fluid ◽  
Nicotiana Tabacum L ◽  
The One

Intercellular fluid b proteins from hypersensitive Nicotiana tabacum L. cv. Xanthi-nc and N. sylvestris Speg. and Comes infected with tobacco mosaic virus were compared by two-dimensional (2-D) polyacrylamide gel electrophoresis. Except for missing bands b2, b6a, b6b, and b7b, the overall 2-D electrophoretic pattern of N. sylvestris intercellular fluid proteins was similar to the one observed with 'Xanthi-nc' tobacco. Intercellular proteins were also studied by chromatography on con-canavalin A. Glycoproteins corresponding to b6a and b7a proteins of N. tabacum and the [Formula: see text] analog of N. sylvestris were identified. These proteins are probably peroxidase isozymes, as peroxidase activities with the same electrophoretic mobility were detected after polyacrylamide gel electrophoresis. No esterase activity was associated with any b protein band in gels. Esterase activities decreased upon virus infection, but accumulation of b proteins and peroxidase activities increased.

scholarly journals Allamanda Mosaic Caused by Cucumber mosaic virus in Taiwan

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 529-529 ◽  
Author(s):  
Y. K. Chen ◽  
C. C. Yang ◽  
H. T. Hsu
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Chenopodium Quinoa ◽  
Rabbit Antiserum ◽  
Nucleotide Sequence Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Indicator Plants

Allamanda (Allamanda cathartica L., family Apocynaceae) is native to Brazil and is a popular perennial shrub or vine ornamental in Taiwan. Plants showing severe mosaic, rugosity, and leaf distortion symptoms on leaves are common in commercial nurseries and private gardens. Examination of crude sap prepared from symptomatic leaves using an electron microscope revealed the presence of spherical virus particles with a diameter of approximately 28 nm. The virus was mechanically transmitted to indicator plants and induced symptoms similar to those incited by Cucumber mosaic virus (CMV). The virus caused local lesions on inoculated leaves of Chenopodium quinoa and C. amaranticolor and systemic mosaic in Cucumis sativus, Lycopersicon esculentum, Nicotiana benthamiana, N. glutinosa, N. rustica, and N. tabacum. On N. tabacum, necrotic ringspots developed on inoculated leaves followed by systemic mosaic. Tests of leaf sap extracted from naturally infected allamanda and inoculated indicator plants using enzyme-linked immunosorbent assay were positive to rabbit antiserum prepared to CMV. Viral coat protein on transblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis reacted with CMV subgroup I specific monoclonal antibodies (2). With primers specific to the 3′-half of RNA 3 (1), amplicons of an expected size (1,115 bp) were obtained in reverse transcription-polymerase chain reaction (RT-PCR) using total RNA extracted from infected allamanda and N. benthamiana. The amplified fragment (EMBL Accession No. AJ871492) was cloned and sequenced. It encompasses the 3′ part of the intergenic region of RNA 3 (158 nt), CP ORF (657 nt), and 3′ NTR (300 nt) showing 91.8–98.9% and 71.4–72.8% identities to those of CMV in subgroups I and II, respectively. Results of MspI-digested restriction fragment length polymorphism patterns of the RT-PCR fragment and the nucleotide sequence analysis indicate that the CMV isolate from allamanda belongs to subgroup IB, which is predominant on the island. To our knowledge, CMV is the only reported virus that infects allamanda and was first detected in Brazil (3), and this is the first report of CMV infection in allamanda plants occurring in Taiwan. References: (1) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (2) H. T. Hsu et al. Phytopathology 90:615, 2000. (3) E. W. Kitajima. Acta. Hortic. 234:451, 1988.

Structural polypeptides of cholera bacteriophage

1981 ◽  
Vol 27 (1) ◽  
pp. 72-75 ◽  
Author(s):  
K. Chaudhuri ◽  
M. Maiti
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Total Protein ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Molecular Weights

The structural polypeptides of the cholera bacteriophage [Formula: see text] have been analysed by sodium dodecyl sulphate – polyacrylamide gel electrophoresis. Eight different polypeptides were identified. The apparent molecular weights of the polypeptides were 143 000, 96 500, 68 000, 53 000, 37 500, 29 500, 21 000, and 13 500, respectively. The percentage of total protein corresponding to each polypeptide was estimated.

scholarly journals Incidencia y sintomatología de cinco virus en parcelas comerciales de chile seco en Aguascalientes, San Luis Potosí y Zacatecas, México

2018 ◽  
Vol 3 (2) ◽  
pp. 381-390
Author(s):  
Rodolfo Velásquez-Valle ◽  
Luis Roberto Reveles-Torres ◽  
Jaime Mena-Covarrubias
Keyword(s):  
Tobacco Mosaic Virus ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Tobacco Etch Virus ◽  
Mottle Virus ◽  
Pepper Mottle Virus ◽  
San Luis ◽  
Das Elisa

A nivel mundial el cultivo de chile es afectado por más de 60 enfermedades virales; sin embargo, poco se conoce acerca de ellas en el área productora de chile seco del norte centro de México por lo que el objetivo del presente trabajo consistió en detectar la presencia y sintomatología de cinco virus en parcelas comerciales de chile seco en los estados mencionados. Plantas de chile de los tipos mirasol y ancho fueron muestreadas y se anotó la presencia de síntomas como enanismo, clorosis, deformación de hojas, defoliación, necrosis vascular y ramas unidas. Las muestras fueron analizadas mediante la técnica DAS- ELISA empleando los antisueros para el virus del mosaico del tabaco (Tobacco mosaic virus: TMV), mosaico del pepino (Cucumber mosaic virus: CMV), Y de la papa (Potato virus Y: PVY), moteado del chile (Pepper mottle virus: PepMoV) y jaspeado del tabaco (Tobacco etch virus: TEV). Esos virus fueron identificados en plantas de chile colectadas en las parcelas comerciales de chile seco de los tres estados antes mencionados.

scholarly journals Molecular Detection of Cucumber Mosaic Virus and Tobacco Mosaic Virus Infecting African Nightshades (Solanum scabrum Miller)

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
S. L. Kimaru ◽  
D. C. Kilalo ◽  
W. M. Muiru ◽  
J. W. Kimenju ◽  
C. R. Thuku
Keyword(s):  
Tobacco Mosaic Virus ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Illumina Miseq ◽  
Amino Acid Sequences ◽  
Management Options ◽  
Phylogenetic Studies ◽  
The Usa ◽  
Miseq Platform ◽  
Clade 1

The two viruses cucumber mosaic virus (CMV) and tobacco mosaic virus (TMV) are among the major viruses that constrain the production of African nightshade (ANS). The ANS provides high proportions of micronutrient content and has medicinal, economic, and agronomic benefits. In this study, we utilized molecular methods to detect these viruses present in ANS at two locations in Kenya: Kisii and Kakamega counties. Ribonucleic acid (RNA) was extracted from leaves of ANS plants exhibiting viral symptoms and sequenced on the Illumina MiSeq platform for phylogenetic studies. The isolates were grouped based on nucleotide and sequence identity. We detected two isolates of CMV and one isolate of TMV in ANS samples. The isolate sequences have been deposited in the GeneBank to obtain accession numbers. Cucumber mosaic virus nucleotide sequence closely resembled the Kirinyaga isolates in Kenya classified in subgroup I. Tobacco mosaic virus phylogenetic studies revealed close resemblance of isolates KY810785.1 from Britain and AF273221.1 from the USA in clade 1 based on nucleotide and amino acid sequences. This is an indication that the virus is widely distributed across the world. Detection of these two viruses in Kenya suggests that they are prevalent in crop-growing regions and the germplasms. African nightshade could also act as a virus reservoir infecting other plants. This study will inform management options to prevent virus epidemics and be in control of vectors.

PROFIL PROTEIN VAKSIN Aeromonas hydrophila DAN Streptococcus agalactiae HASIL INAKTIVASI DENGAN FORMALIN: DIUJI MENGGUNAKAN Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis

2014 ◽  
Vol 9 (3) ◽  
pp. 449
Author(s):  
Desy Sugiani ◽  
Angela Mariana Lusiastuti ◽  
Sukenda Sukenda ◽  
Enang Harris
Keyword(s):  
Streptococcus Agalactiae ◽  
Sodium Dodecyl Sulphate ◽  
Total Protein ◽  
Aeromonas Hydrophila ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Sds Page ◽  
Neutral Buffer Formalin

Vaksin bakterin dalam bentuk protein merupakan salah satu tipe vaksin yang telah banyak dikembangkan. Protein digunakan sebagai vaksin biasanya dibuat dengan teknik inaktivasi formalin-killed. Vaksin ini biasanya lebih mudah dibuat, lebih murah, lebih stabil, dan mampu disimpan dalam waktu lama. Akan tetapi masih sedikit informasi mengenai efek perlakuan tersebut terhadap profil protein. Pada penelitian ini, untuk mengevaluasi profil protein, dilakukan inaktivasi sediaan vaksin dari isolat bakteri Aeromonas hydrophila AHL0905-2 dan Streptococcus agalactiae N14G dengan menambahkan 0,5% formalin dan 3% neutral buffer formalin (NBF) ke dalam biakan plasebo bakterin dan diinkubasi selama 24 jam. Kualitas produk vaksin ditentukan berdasarkan uji karakterisasi protein menggunakan metode Bradford dan SDS-PAGE. Hasil uji menunjukkan bahwa sediaan vaksin A. hydrophila dan S. agalactiae yang diinaktivasi dengan 3% NBF memiliki profil protein lebih variatif dibandingkan dengan sediaan vaksin yang diinaktivasi dengan 0,5% formalin. Akan tetapi, inaktivasi vaksin A. hydrophila dan S. agalactiae dengan 3% NBF menghasilkan berat total protein yang lebih rendah jika dibandingkan dengan dengan sediaan vaksin yang diinaktivasi dengan 0,5% formalin.

Protein Profile of Human Perilymph: In Search of Markers for the Diagnosis of Perilymph Fistula and other Inner Ear Disease

1994 ◽  
Vol 111 (3P1) ◽  
pp. 273-280 ◽  
Author(s):  
Isolde Thalmann ◽  
Robert I. Kohut ◽  
Jay Ryu ◽  
Thomas H. Comegys ◽  
Masamitsu Senarita ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Inner Ear ◽  
Total Protein ◽  
Gel Electrophoresis ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Density Lipoprotein ◽  
Polyacrylamide Gel ◽  
Computer Assisted ◽  
Two Dimensional

Recent developments in high-resolution two-dimensional polyacrylamide gel electrophoresis, combined with amino acid sequencing and computer-assisted image analysis, have allowed separation of approximately 100 proteins and identification and quantitation of some 30 proteins in human perilymph. The majority of proteins were found to be present in perilymph at levels in basic agreement with the total protein gradient between perilymph and plasma (1:35). However, several striking differences were observed: (1) β2-transferrin, known to be absent from normal plasma but present in cerebrospinal fluid, was detected in perilymph at a concentration roughly equal to that in cerebrospinal fluid; and (2) two high-density lipoprotein-associated apolipoproteins—apo D (formerly PLS:33) and apo J or NA1 and NA2 (formerly PSL:29/30), the latter showing identity with 5P40/40, or cytolysis inhibitor—were found to be present at concentrations 1 to 2 orders of magnitude higher when examined in terms of total protein and to be comparable with or higher than plasma levels when examined in terms of absolute concentrations. The functional significance of the extremely high levels of the two apolipoproteins is not known at this time. An attempt was made to use β2-transferrin, as well as apo D and apo J (NA1/NA2), as markers for the diagnosis of perilymph fistula, one of the most controversial and challenging problems for the otologist today. It was determined that the technique is indeed applicable when relatively pure fistula samples are analyzed. Limitations and potential improvements of the technique are discussed. In addition, the potential usefulness of two-dimensional polyacrylamide gel electrophoresis in other pathologic conditions of the inner ear is discussed briefly.

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