scholarly journals Coupling Virus-Induced Gene Silencing to Exogenous Green Fluorescence Protein Expression Provides a Highly Efficient System for Functional Genomics in Arabidopsis and across All Stages of Tomato Fruit Development

2011 ◽  
Vol 156 (3) ◽  
pp. 1278-1291 ◽  
Author(s):  
Leandro Quadrana ◽  
Maria Cecilia Rodriguez ◽  
Mariana López ◽  
Luisa Bermúdez ◽  
Adriano Nunes-Nesi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Gene Silencing ◽  
Fruit Development ◽  
Tomato Fruit ◽  
Green Fluorescence Protein ◽  
Virus Induced Gene Silencing ◽  
Green Fluorescence ◽  
Efficient System ◽  
Green Fluorescence Protein Expression ◽  
Fluorescence Protein

scholarly journals Expansion of EasyClone-MarkerFree toolkit for Saccharomyces cerevisiae genome with new integration sites

2021 ◽  
Author(s):  
Mahsa Babaei ◽  
Luisa Sartori ◽  
Alexey Karpukhin ◽  
Dmitrii Abashkin ◽  
Elena Matrosova ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Recombinant Dna ◽  
Green Fluorescence Protein ◽  
Cellular Growth ◽  
Green Fluorescence ◽  
Biotechnological Production ◽  
Yeast Saccharomyces Cerevisiae ◽  
Integration Sites ◽  
Fluorescence Protein ◽  
Integration Efficiency

Abstract Biotechnological production requires genetically stable recombinant strains. To ensure genomic stability, recombinant DNA is commonly integrated into the genome of the host strain. Multiple genetic tools have been developed for genomic integration into baker's yeast Saccharomyces cerevisiae. Previously, we had developed a vector toolkit EasyClone-MarkerFree for stable integration into eleven sites on chromosomes X, XI, and XII of S. cerevisiae. The markerless integration was enabled by CRISPR-Cas9 system. In this study, we have expanded the kit with eight additional intergenic integration sites located on different chromosomes. The integration efficiency into the new sites was above 80%. The expression level of green fluorescence protein (gfp) for all eight sites was similar or above XI-2 site from the original EasyClone-MarkerFree toolkit. The cellular growth was not affected by the integration into any of the new eight locations. The eight-vector expansion kit is available from AddGene.

Establishment of an Embryotoxicity Assay with Green Fluorescence Protein-expressing Embryonic Cell-derived Cardiomyocytes

1999 ◽  
Vol 27 (3) ◽  
pp. 471-484 ◽  
Author(s):  
Susanne Bremer ◽  
Maaike Van Dooren ◽  
Martin Paparella ◽  
Eugen Kossolov ◽  
Bernd Fleischmann ◽  
...  
Keyword(s):  
Green Fluorescence Protein ◽  
Embryonic Cell ◽  
Green Fluorescence ◽  
Fluorescence Protein

scholarly journals Fertility of mice receiving vitrified adult mouse ovaries

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 681-687 ◽  
Author(s):  
Toshio Hani ◽  
Takanori Tachibe ◽  
Saburo Shingai ◽  
Nobuo Kamada ◽  
Otoya Ueda ◽  
...  
Keyword(s):  
Germ Cells ◽  
Adult Mouse ◽  
Green Fluorescence Protein ◽  
Green Fluorescence ◽  
Experimental Animals ◽  
Immature Mouse ◽  
Estrous Cyclicity ◽  
Fluorescence Protein ◽  
High Degree ◽  
Old Mice

Cryopreservation of the ovaries is a useful technology for preservation of germ cells from experimental animals, because if the female founder is infertile or has mutated mitochondrial DNA, preservation of female germ cells is necessary. Although it is possible to cryopreserve immature mouse ovaries with a high degree of viability by vitrification with a mixture of several cryoprotectants, the viability of cryopreserved adult mouse ovaries is still unknown. Here, we investigated the viability of mouse ovaries at various ages after cryopreservation by vitrification techniques. Donor ovaries were collected from 10-day-, 4-week-, 10-week- and 7-month-old, female, nulliparous, green fluorescence protein (GFP)-transgenic mice and cryopreserved by vitrification. The vitrified-warmed ovaries were orthotopically transplanted to 4- or 10-week-old mice. GFP-positive pups were obtained in all experimental groups. In the 4-week-old recipients, the percentages of GFP-positive pups among the total pups from recipients transplanted with ovaries of 10-day-, 4-week-, 10-week- and 7-month-old donors were 44%, 9%, 12% and 4% respectively. In the 10-week-old recipients, the percentages of GFP-positive pups among the total pups from recipients transplanted with ovaries of 10-day-, 4-week-, 10-week- and 7-month-old donors were 36%, 16%, 2% and 9% respectively. Furthermore, GFP-positive pups also were obtained from recipients transplanted with ovaries of donors without normal estrous cyclicity. Our results indicate that cryopreservation of mouse ovaries by vitrification is a useful method for the preservation of female germ cells from mice of various ages.

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