scholarly journals Regeneration and transformation of Polish cultivars of potato

2014 ◽  
Vol 64 (4) ◽  
pp. 335-340
Author(s):  
Anna Nadolska-Orczyk ◽  
Lidia Miłkowska ◽  
Andrzej Pałucha ◽  
Paweł Czembor ◽  
Wacław Orczyk
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Transgenic Plants ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Transformation Efficiency ◽  
Binary Vector ◽  
Leaf Explants ◽  
Vector System ◽  
Regeneration Ability

The article presents the results of regeneration and transformation experiments of 12 Polish cultivars of potato. The cultivars Brda, Bzura, Elipsa and Irga regenerated the highest number of shoots from leaf explants (60% to 100% of explants regenerated 4.9 to 16.5 shoots per explant). The cultivars Brda, Bzura, Elipsa and Irys were the source of the best regenerating tuber explants (66% to 100% of explants regenerated 6.2 to 11.9 shoots from one explant). Both types of explants (from leaves and tubers) were used for transformation experiments. <i>Agrobacterium tumefaciens</i> strains used for transformation contained binary vector system: LBA 4404 (pAL 4404:pBI 121) and C58C1 (pGV2260:pVU104). There was a strong correlation between regeneration ability of tested cultivars and transformation efficiency. GUS-positive, kanamycin resistant and well rooted plants from leaf explants in cultivars Bzura, Brda and from tuber explants in Bzura and Elipsa were obtained. Northern blot analysis confirmed the presence of β-glucuronidase mRNA in transgenic plants.

Genetic Modification of `Royal Gala' Apple by Agrobacterium-mediated Transformation and Colchicine-induced Mutation

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461b-461
Author(s):  
Qingzhong Liu ◽  
Freddi Hammerschlag ◽  
Rengong Meng
Keyword(s):  
Flow Cytometry ◽  
Disease Resistance ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Transformation Efficiency ◽  
Binary Vector ◽  
Leaf Explants ◽  
Wild Type ◽  
Regeneration Medium ◽  
Osmotin Promoter

As part of a program to develop transgenic Malus × domestica Borkh. cv. Royal Gala with improved disease resistance, transgenic diploid and tetraploid plants with cecropin MB39 gene were regenerated. Transgenic diploid plants were obtained from etiolated internodal explants by Agrobacterium tumefaciens-mediated transformation using the plasmid binary vector pGV containing a chimeric gene consisting of a secretory sequence from barley-amylase joined to a modified cecropin MB39 coding sequence and placed under control of wound-inducible osmotin promoter from tobacco. The integration of the cecropin gene into apple genome was confirmed by Southern blot analysis. The transformation efficiency was 1.5%. Both non- and transgenic tetraploid plants were produced by cocultivating leaf explants from wild type and transgenic diploid shoots with colchicine at 25 mg/L in apple regeneration medium containing 10 μM TDZ. Twenty-two tetraploid lines were obtained from 90 explants. Flow cytometry was used for ploidy determination. The tetraploid plants were distinguishable from the diploid on morphological as well as cytogenetic grounds. Both the transgenic diploid and tetraploid plants are now being evaluated for resistance to fireblight.

scholarly journals IL-8 and MCP Gene Expression and Production by LPS-Stimulated Human Corneal Stromal Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
Roni M. Shtein ◽  
Susan G. Elner ◽  
Zong-Mei Bian ◽  
Victor M. Elner
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Stromal Cells ◽  
Blot Analysis ◽  
Time Course ◽  
Northern Blot Analysis ◽  
Statistical Significance ◽  
Dependent Manner ◽  
Corneal Inflammation ◽  
Chemotactic Protein

Purpose. To determine time course of effect of lipopolysaccharide (LPS) on production of interleukin-8 (IL-8) and monocyte chemotactic protein (MCP) by cultured human corneal stromal cells.Methods. Human corneal stromal cells were harvested from donor corneal specimens, and fourth to sixth passaged cells were used. Cell cultures were stimulated with LPS for 2, 4, 8, and 24 hours. Northern blot analysis of IL-8 and MCP gene expression and ELISA for IL-8 and MCP secretion were performed. ELISA results were analyzed for statistical significance using two-tailed Student'st-test.Results. Northern blot analysis demonstrated significantly increased IL-8 and MCP gene expression after 4 and 8 hours of exposure to LPS. ELISA for secreted IL-8 and MCP demonstrated statistically significant increases (P<0.05) after corneal stromal cell stimulation with LPS.Conclusions. This paper suggests that human corneal stromal cells may participate in corneal inflammation by secreting potent leukocyte chemotactic and activating proteins in a time-dependent manner when exposed to LPS.

scholarly journals A rapid and accurate method for quantitating total RNA transferred during Northern blot analysis

1988 ◽  
Vol 16 (5) ◽  
pp. 2354-2354 ◽  
Author(s):  
Nathalie Denis ◽  
Daniel Corcos ◽  
Jacques Kruh ◽  
Alain Kitzis
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Accurate Method ◽  
Total Rna

The mouse prostaglandin E receptor EP2 subtype: cloning, expression, and Northern blot analysis

FEBS Letters ◽  
1995 ◽  
Vol 372 (2-3) ◽  
pp. 151-156 ◽  
Author(s):  
Masato Katsuyama ◽  
Nobuhiro Nishigaki ◽  
Yukihiko Sugimoto ◽  
Kimiko Morimoto ◽  
Manabu Negishi ◽  
...  
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Prostaglandin E

A procedure for Northern blot analysis of native RNA

1986 ◽  
Vol 159 (1) ◽  
pp. 227-232 ◽  
Author(s):  
Edouard W. Khandjian ◽  
Claude Méric
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis

The expression of NG2 proteoglycan in the developing rat limb

Development ◽  
1991 ◽  
Vol 111 (4) ◽  
pp. 933-944 ◽  
Author(s):  
A. Nishiyama ◽  
K.J. Dahlin ◽  
W.B. Stallcup
Keyword(s):  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Core Protein ◽  
Staining Intensity ◽  
Limb Bud ◽  
Protein Size ◽  
Glial Progenitor Cells ◽  
Ng2 Proteoglycan ◽  
Developing Rat

NG2 is a chondroitin sulfate proteoglycan previously found to be expressed by glial progenitor cells of the O2A lineage. We have examined the expression of NG2 in the developing rat limb by immunohistochemistry and northern blot analysis. Staining of embryonic day 14 (E14) rat limb bud sections with polyclonal and monoclonal anti-NG2 antibodies reveals reactivity in the precartilaginous mesenchymal condensation. The staining intensity increases with the differentiation of chondrocytes until E16. NG2 staining is not detected in the mature hypertrophic chondrocytes of E17 and postnatal day 3 (P3) limbs even after treatment of the sections with hyaluronidase or collagenase. Immuno-precipitations with anti-NG2 antibody using 125I-labeled limb cells in culture showed a 400 to 800 × 10(3) Mr proteoglycan species with a core protein size of 300 × 10(3) Mr, comparable to NG2 from O2A cells and neural cell lines. Northern blot analysis reveals the expression of an 8.9 kb mRNA in E16 limbs and at a lower level in P1 cartilage. The northern blot analyses also show that NG2 is distinct from the large aggregating proteoglycan of the cartilage. Our results indicate that in the developing limb cartilage, as in the differentiating oligodendrocytes, NG2 is present on immature cells in the process of differentiating, but its expression is downregulated as terminal differentiation of chondrocytes takes place.

PKC-lambda is the atypical protein kinase C isoform expressed by immature ventricle

1997 ◽  
Vol 272 (4) ◽  
pp. H1636-H1642
Author(s):  
V. O. Rybin ◽  
P. M. Buttrick ◽  
S. F. Steinberg
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Ventricular Myocardium ◽  
Adult Rat ◽  
Atypical Pkc ◽  
Kinase C ◽  
Pkc Zeta

We recently identified a developmental decline in protein kinase C (PKC) isoform expression, at the level of the protein, in rat ventricular myocardium. To investigate mechanisms regulating PKC isoform expression in cardiac tissue, this study uses Northern blot analysis to compare the abundance of PKC isoform mRNAs in neonatal and adult rat ventricular myocardium. PKC-epsilon protein and mRNA were detected in both neonatal and adult rat ventricular myocardial preparations. In contrast, coordinate postnatal declines in the abundance of PKC-alpha and PKC-delta proteins and transcripts were identified. An antiserum raised against the COOH-terminal sequence of PKC-zeta detected abundant immunoreactivity in neonatal, but not adult, ventricular myocytes. However, PKC-zeta transcripts were not detectable in the heart either by Northern blot analysis or a reverse transcriptase-polymerase chain reaction approach, indicating that neither the myocytes nor the contaminating cellular elements in the heart express PKC-zeta. Rather, PKC-lambda, another atypical PKC isoform that is structurally highly homologous to PKC-zeta, was detected at the protein and mRNA level in neonatal, but not adult, ventricular myocardium. Taken together, these results establish that developmental declines in calcium-sensitive, novel, and atypical PKC isoforms are paralleled by changes in the levels of the mRNAs encoding these proteins, suggesting transcriptional regulation of PKC during normal cardiac development. The results of this study further identify PKC-lambda as the atypical PKC isoform expressed by the immature ventricle.

Sign in / Sign up

Export Citation Format

Share Document