Tissue culture of wild-type, interspecific genome/plastome hybrids and plastome mutants of evening primrose ( Oenothera) : controlled morphogenesis and transformation

1998 ◽  
Vol 17 (8) ◽  
pp. 605-611 ◽  
Author(s):  
A. Mehra-Palta ◽  
H.-U. Koop ◽  
S. Goes ◽  
E.-M. Troidl ◽  
G. Nagy ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Wild Type ◽  
Evening Primrose ◽  
Plastome Mutants

scholarly journals Quantification of Hepatitis A Virus in Shellfish by Competitive Reverse Transcription-PCR with Coextraction of Standard RNA

1999 ◽  
Vol 65 (1) ◽  
pp. 322-326 ◽  
Author(s):  
Charlotte Arnal ◽  
Virginie Ferre-Aubineau ◽  
Berangere Mignotte ◽  
Berthe Marie Imbert-Marcille ◽  
Sylviane Billaudel
Keyword(s):  
Tissue Culture ◽  
Reverse Transcription ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Internal Standard ◽  
Wild Type ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Reproducible Method ◽  
Dna Enzyme Immunoassay

ABSTRACT To quantify hepatitis A virus (HAV) in experimentally contaminated mussels, we developed an internal standard RNA with a 7-nucleotide deletion for competitive reverse transcription (RT)-PCR. Deposited directly into the sample, this standard was used both as extraction control and as quantification tool. After coextraction and competitive RT-PCR, standard and wild-type products were detected by differential hybridization with specific probes and a DNA enzyme immunoassay. The quantifiable range with this reproducible method was 104 to 107 copies of HAV/gram or 400 to 106 50% tissue culture infective doses/ml.

Enhancing Biosynthesis of Flavonol Protective Biomaterials Using FLS Transgenic Plants

2017 ◽  
Vol 866 ◽  
pp. 29-32
Author(s):  
Darin Dangrit ◽  
Kanokporn Sompornpailin
Keyword(s):  
Tissue Culture ◽  
Transgenic Plants ◽  
Photosynthetic Pigment ◽  
Flavonoid Biosynthesis ◽  
Carotenoid Pigments ◽  
Chlorophyll B ◽  
Wild Type ◽  
Flavonoid Synthesis ◽  
Transgenic Lines ◽  
Tissue Culture Condition

Flavonol synthase (FLS) gene encodes an enzyme that is involved in conversion substrates into flavonols, quercetin and kaempferol. These substances are a subgroup of flavonoids which have an important role in both plant and human health. Many environmental factors such as temperature, pH and UV-A radiation have been studied and presented relationship with flavonoid synthesis. In this experiment, the combination of visible and UV-A lights was used as factors for elevating flavonoid biosynthesis of wild type (WT) plant and two lines of FLS transgenic plant under tissue culture condition. Both transgenic lines significantly enhanced the accumulation of quercetin and kaempferol substances nearly one fold higher than WT plant did. The photosynthetic pigment levels of chlorophyll A, chlorophyll B and carotenoid in transgenic lines are in the range 45.20-46.88, 16.34-17.04 and 13.63-13.46, while those of WT plants are 35.93, 13.18 and 10.55 (µg/g FW), respectively. Therefore, FLS transgenic plants containing high flavonol content showed a better in the protection photosynthetic pigments by less reductions of chlorophyll and carotenoid pigments.

scholarly journals Complete Genome Sequences of SevenHelicoverpa armigeraSNPV-AC53-Derived Strains

2016 ◽  
Vol 4 (3) ◽  
Author(s):  
Christopher Noune ◽  
Caroline Hauxwell
Keyword(s):  
Tissue Culture ◽  
Helicoverpa Armigera ◽  
Complete Genome ◽  
Full Genome ◽  
Wild Type ◽  
Genome Sequences ◽  
Helicoverpa Armígera ◽  
Multiple Strains

Wild-type baculovirus isolates typically consist of multiple strains. We report the full genome sequences of seven alphabaculovirus strains derived by passage through tissue culture fromHelicoverpa armigeraSNPV-AC53 (KJ909666).

scholarly journals Cloning and expression of two ornithine decarboxylase forms from HMOA cells

1995 ◽  
Vol 312 (1) ◽  
pp. 13-16
Author(s):  
R Autelli ◽  
L Persson ◽  
F M Baccino
Keyword(s):  
Tissue Culture ◽  
Ornithine Decarboxylase ◽  
Half Life ◽  
Turnover Rate ◽  
Cloning And Expression ◽  
Single Amino Acid ◽  
Turnover Rates ◽  
Wild Type ◽  
First Time ◽  
Hepatoma Tissue

In HMOA cells [Mamont, Duchesne, Grove and Tardif (1978) Exp. Cell Res. 115, 387-393] the half-life of ornithine decarboxylase (ODC) is 8-14 h instead of 15 min as in the Hepatoma Tissue Culture parental cells, due to a single amino acid substitution [Miyazaki, Matsufuji, Murakami and Hayashi (1993) Eur. J. Biochem. 214, 837-844]. We demonstrate for the first time that HMOA cells possess two forms of ODC mRNA that are translated into two proteins differing greatly in turnover rates. We have cloned and transfected the cDNAs for the two ODC forms into COS-1 cells for a direct measurement of their turnover rate. The variant ODC form was much more stable than the wild-type protein, with a half-life of 14 h as compared with 2.5 h.

scholarly journals A Hypervariable Region within the 3′ cis-Acting Element of the Murine Coronavirus Genome Is Nonessential for RNA Synthesis but Affects Pathogenesis

2006 ◽  
Vol 81 (3) ◽  
pp. 1274-1287 ◽  
Author(s):  
Scott J. Goebel ◽  
Timothy B. Miller ◽  
Corey J. Bennett ◽  
Kristen A. Bernard ◽  
Paul S. Masters
Keyword(s):  
Tissue Culture ◽  
Deletion Mutant ◽  
Rna Synthesis ◽  
Mutational Analysis ◽  
Hepatitis Virus ◽  
Point Mutations ◽  
Hypervariable Region ◽  
Wild Type Virus ◽  
Wild Type ◽  
Group 2

ABSTRACT The 3′ cis-acting element for mouse hepatitis virus (MHV) RNA synthesis resides entirely within the 301-nucleotide 3′ untranslated region (3′ UTR) of the viral genome and consists of three regions. Encompassing the upstream end of the 3′ UTR are a bulged stem-loop and an overlapping RNA pseudoknot, both of which are essential to MHV and common to all group 2 coronaviruses. At the downstream end of the genome is the minimal signal for initiation of negative-strand RNA synthesis. Between these two ends is a hypervariable region (HVR) that is only poorly conserved between MHV and other group 2 coronaviruses. Paradoxically, buried within the HVR is an octanucleotide motif (oct), 5′-GGAAGAGC-3′, which is almost universally conserved in coronaviruses and is therefore assumed to have a critical biological function. We conducted an extensive mutational analysis of the HVR. Surprisingly, this region tolerated numerous deletions, rearrangements, and point mutations. Most striking, a mutant deleted of the entire HVR was only minimally impaired in tissue culture relative to the wild type. By contrast, the HVR deletion mutant was highly attenuated in mice, causing no signs of clinical disease and minimal weight loss compared to wild-type virus. Correspondingly, replication of the HVR deletion mutant in the brains of mice was greatly reduced compared to that of the wild type. Our results show that neither the HVR nor oct is essential for the basic mechanism of MHV RNA synthesis in tissue culture. However, the HVR appears to play a significant role in viral pathogenesis.

scholarly journals A Double-Selective Tissue Culture System for Isolation of Wild-Type Poliovirus from Sewage Applied in a Long-Term Environmental Surveillance

1999 ◽  
Vol 65 (4) ◽  
pp. 1794-1797 ◽  
Author(s):  
Y. Manor ◽  
R. Handsher ◽  
T. Halmut ◽  
M. Neuman ◽  
B. Abramovitz ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Detection Limit ◽  
Culture System ◽  
Environmental Surveillance ◽  
Wild Type ◽  
Selective Method ◽  
Tissue Culture System ◽  
Cost Efficient

ABSTRACT We describe a simple, cost-efficient, double-selective method for isolation of wild-type poliovirus from sewage samples containing vaccine polioviruses and other enteroviruses, with a detection limit of 18 to 50 PFU per 1 to 2 liters of sewage. By this method we were able to process 1,700 sewage samples collected between 1991 and 1996, from which 10,472 plaques were isolated, 41 of them being identified as wild-type polioviruses.

scholarly journals Genetic Determinants of Japanese Encephalitis Virus Vaccine Strain SA14-14-2 That Govern Attenuation of Virulence in Mice

2015 ◽  
Vol 89 (12) ◽  
pp. 6328-6337 ◽  
Author(s):  
Gregory D. Gromowski ◽  
Cai-Yen Firestone ◽  
Stephen S. Whitehead
Keyword(s):  
Tissue Culture ◽  
Amino Acid ◽  
Japanese Encephalitis Virus ◽  
Vaccine Strain ◽  
Japanese Encephalitis ◽  
Second Generation ◽  
Virus Genome ◽  
Vaccine Virus ◽  
Wild Type ◽  
Virulence Attenuation

ABSTRACTThe safety and efficacy of the live-attenuated Japanese encephalitis virus (JEV) SA14-14-2 vaccine are attributed to mutations that accumulated in the viral genome during its derivation. However, little is known about the contribution that is made by most of these mutations to virulence attenuation and vaccine immunogenicity. Here, we generated recombinant JEV (rJEV) strains containing JEV SA14-14-2 vaccine-specific mutations that are located in the untranslated regions (UTRs) and seven protein genes or are introduced from PCR-amplified regions of the JEV SA14-14-2 genome. The resulting mutant viruses were evaluated in tissue culture and in mice. The authentic JEV SA14-14-2 (E) protein, with amino acid substitutions L107F, E138K, I176V, T177A, E244G, Q264H, K279M, A315V, S366A, and K439R relative to the wild-type rJEV clone, was essential and sufficient for complete attenuation of neurovirulence. Individually, the nucleotide substitution T39A in the 5′ UTR (5′-UTR-T39A), the capsid (C) protein amino acid substitution L66S (C-L66S), and the complete NS1/2A genome region containing 10 mutations each significantly reduced virus neuroinvasion but not neurovirulence. The levels of peripheral virulence attenuation imposed by the 5′-UTR-T39A and C-L66S mutations, individually, were somewhat mitigated in combination with other vaccine strain-specific mutations, which might be compensatory, and together did not affect immunogenicity. However, a marked reduction in immunogenicity was observed with the addition of the NS1/2A and NS5 vaccine virus genome regions. These results suggest that a second-generation recombinant vaccine can be rationally engineered to maximize levels of immunogenicity without compromising safety.IMPORTANCEThe live-attenuated JEV SA14-14-2 vaccine has been vital for controlling the incidence of disease caused by JEV, particularly in rural areas of Asia where it is endemic. The vaccine was developed >25 years ago by passaging wild-type JEV strain SA14 in tissue cultures and rodents, with intermittent tissue culture plaque purifications, to produce a virus clone that had adequate levels of attenuation and immunogenicity. The vaccine and parent virus sequences were later compared, and mutations were identified throughout the vaccine virus genome, but their contributions to attenuation were never fully elucidated. Here, using reverse genetics, we comprehensively defined the impact of JEV SA14-14-2 mutations on attenuation of virulence and immunogenicity in mice. These results are relevant for quality control of new lots of the current live-attenuated vaccine and provide insight for the rational design of second-generation, live-attenuated, recombinant JEV vaccine candidates.

A filamentous-like mutant of Listeria monocytogenes with reduced expression of a 60-kilodalton extracellular protein invades and grows in 3T6 and Caco-2 cells

1992 ◽  
Vol 38 (8) ◽  
pp. 843-851 ◽  
Author(s):  
Karen A. Gutekunst ◽  
Leo Pine ◽  
Elizabeth White ◽  
Sophia Kathariou ◽  
George M. Carlone
Keyword(s):  
Tissue Culture ◽  
Listeria Monocytogenes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Listeriolysin O ◽  
Wild Type ◽  
Phagocytic Cells ◽  
Electron Microscopic ◽  
Rough Mutant

We describe a spontaneous rough mutant of Listeria monocytogenes that produces reduced amounts of a 60-kilodalton major extracellular polypeptide (p60) as shown by sodium dodecyl sulfate – polyacrylamide gel electrophoresis and Western blot analysis. The cells of this mutant are filamentous, do not give rise to smooth wild-type colonies, and produce listeriolysin O in amounts equal to that of the wild-type cells, but they show a reduced virulence in the mouse LD50 model and in the Caco-2 tissue culture virulence assay. Light and electron microscopic studies show that this mutant invades and remains filamentous during in vivo growth in both Caco-2 and 3T6 tissue culture monolayers. The reduced virulence of the rough mutant is not due to the inability of its filamentous forms to invade or to grow in nonprofessional phagocytes since invasion and growth of the smooth wild-type and the rough mutants are comparable in both Caco-2 and 3T6 monolayers. Key words: Listeria monocytogenes, invasion, nonprofessional phagocytic cells, electron microscopy.

Comparative mineral and hormonal analyses of wild type and TLS somaclonal variant derived from oil palm (Elaeis guineensis Jacq. var. tenera) tissue culture

2012 ◽  
Vol 68 (2) ◽  
pp. 313-317 ◽  
Author(s):  
S. H. Habib ◽  
S.-E. Ooi ◽  
Ondřej Novák ◽  
Danuše Tarkowská ◽  
Jakub Rolčík ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oil Palm ◽  
Elaeis Guineensis ◽  
Wild Type ◽  
Somaclonal Variant ◽  
Elaeis Guineensis Jacq

scholarly journals Genetic and Global Epigenetic Modification, Which Determines the Phenotype of Transgenic Rice?

2020 ◽  
Vol 21 (5) ◽  
pp. 1819
Author(s):  
Xiaoru Fan ◽  
Jingguang Chen ◽  
Yufeng Wu ◽  
CheeHow Teo ◽  
Guohua Xu ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Transgenic Plants ◽  
Plant Height ◽  
Epigenetic Modification ◽  
Epigenetic Changes ◽  
Wild Type ◽  
Global Methylation ◽  
Phenotypic Changes ◽  
Wide Range ◽  
Transgenic Lines

Transgenic technologies have been applied to a wide range of biological research. However, information on the potential epigenetic effects of transgenic technology is still lacking. Here, we show that the transgenic process can simultaneously induce both genetic and epigenetic changes in rice. We analyzed genetic, epigenetic, and phenotypic changes in plants subjected to tissue culture regeneration, using transgenic lines expressing the same coding sequence from two different promoters in transgenic lines of two rice cultivars: Wuyunjing7 (WYJ7) and Nipponbare (NP). We determined the expression of OsNAR2.1 in two overexpression lines generated from the two cultivars, and in the RNA interference (RNAi) OsNAR2.1 line in NP. DNA methylation analyses were performed on wild-type cultivars (WYJ7 and NP), regenerated lines (CK, T0 plants), segregation-derived wild-type from pOsNAR2.1-OsNAR2.1 (SDWT), pOsNAR2.1-OsNAR2.1, pUbi-OsNAR2.1, and RNAi lines. Interestingly, we observed global methylation decreased in the T0 regenerated line of WYJ7 (CK-WJY7) and pOsNAR2.1-OsNAR2.1 lines but increased in pUbi-OsNAR2.1 and RNAi lines of NP. Furthermore, the methylation pattern in SDWT returned to the WYJ7 level after four generations. Phenotypic changes were detected in all the generated lines except for SDWT. Global methylation was found to decrease by 13% in pOsNAR2.1-OsNAR2.1 with an increase in plant height of 4.69% compared with WYJ7, and increased by 18% in pUbi-OsNAR2.1 with an increase of 17.36% in plant height compared with NP. This suggests an absence of a necessary link between global methylation and the phenotype of transgenic plants with OsNAR2.1 gene over-expression. However, epigenetic changes can influence phenotype during tissue culture, as seen in the massive methylation in CK-WYJ7, T0 regenerated lines, resulting in decreased plant height compared with the wild-type, in the absence of a transformed gene. We conclude that in the transgenic lines the phenotype is mainly determined by the nature and function of the transgene after four generations of transformation, while the global epigenetic modification is dependent on the genetic background. Our research suggests an innovative insight in explaining the reason behind the occurrence of transgenic plants with random and undesirable phenotypes.

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