scholarly journals Transformation and confirmation of GUS gene expression in Solanum melongena L. of PLR 1 cultivar

2018 ◽  
Vol 7 (4) ◽  
pp. 2185 ◽  
Author(s):  
Vinod Kanna S. ◽  
Jayabalan N.
Keyword(s):  
Gene Expression ◽  
Southern Hybridization ◽  
Marker Gene ◽  
Solanum Melongena ◽  
Negative Control ◽  
Selection Marker ◽  
Gus Gene ◽  
Specific Primers ◽  
Agrobacterium Strain ◽  
Gus Gene Expression

In the present study GUS gene transformation was carried out in eggplant using Agrobacterium strain with pBAL2 vector harboring gus gene and nptII as selection marker gene. The factors which are affecting (enhancing) the frequency of transient gus gene expression are different physical and biochemical variables has been carried out. It is observed that the 4 day precultured explants showed the minimum survival rate in the medium when compared with 2-day co cultivated medium. The explants which had undergone co-cultivation for 4 to 5 days showed GUS activity, the tissues were adversely affected due to the overgrowth of bacteria. The gene specific primers for nptII and gus gene were used for amplification and it has given 680bp and 1.9 kb amplified fragments respectively and recorded. The band was detected in the selected plants, but it was absent from the negative control (non-transformed) plant in the Southern hybridization. Our experiment showed 0.80-1.60 percentage of efficiency in transformation. With a total of 849 infected shoots were undergone confirmation tests which results 9 PCR positives (1.06% efficiency). The Transformant kept in the Environmental Growth Chamber and transferred to field condition subsequently.

Studies of Cytokinin Action and Metabolism Using Tobacco Plants Expressing either the ipt or the GUS Gene Controlled by a Chalcone Synthase Promoter. IIipt and GUS Gene Expression, Cytokinin Levels and Metabolism

1997 ◽  
Vol 24 (5) ◽  
pp. 673 ◽  
Author(s):  
Jian Wang ◽  
D. S. Letham ◽  
Edwina Cornish ◽  
K. Wei ◽  
C. H. Hocart ◽  
...  
Keyword(s):  
Gene Expression ◽  
Chalcone Synthase ◽  
Hormonal Control ◽  
Mature Leaf ◽  
Cytokinin Oxidase ◽  
Endogenous Cytokinin ◽  
Gus Gene ◽  
Ipt Gene ◽  
Cytokinin Level ◽  
Gus Gene Expression

The expression of GUS and ipt genes under control of a chalcone synthase (chs) promoter (PCHS) has been determined in tobacco (Nicotiana tabacum L.) plants and related to the development of plants expressing the chimaeric PCHS -ipt gene. GUS gene expression, which served as a model for the expression of the ipt gene, was highest in the internal phloem tissue of stems, in mature leaf laminae and in the upper part of corollas when fully open. Expression of the PCHS -ipt gene was assessed by quantifying the cytokinins produced, by determining incorporation of [3H]adenine into cytokinins and by quantifying ipt mRNA. Results from these studies were in general agreement with those based on expression of the PCHS -GUS gene. The chs promoter controlled expression of the ipt gene with some degree of tissue and temporal specificity. Expression of the ipt gene markedly elevated the cytokinin level in mature leaf laminae and the upper stems of flowering plants. The former was associated with retardation of leaf senescence and increased rates of transpiration due to changes in number, size and aperture of stomata, while the latter was associated with development of lateral shoots. In shoot tip cultures, 2-fold elevations in endogenous cytokinin level caused clear changes in development and this is discussed in relation to current concepts concerning the hormonal control of plant development. Using the transgenic tobacco tissues, it was shown that cis-zeatin is a substrate for cytokinin oxidase, that cis-zeatin is not converted to trans-zeatin in these tissues and that the endogenous cytokinin level influences the level of cytokinin oxidase activity in tissue and the rate of degradation of exogenous zeatin riboside to adenosine.

scholarly journals CRISPR/Transposon gene integration (CRITGI) can manage gene expression in a retrotransposon-dependent manner

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Miki Hanasaki ◽  
Hiroshi Masumoto
Keyword(s):  
Gene Expression ◽  
Amino Acid ◽  
Consensus Sequence ◽  
Marker Gene ◽  
Gene Activation ◽  
Fine Tuning ◽  
Selection Marker ◽  
Regulation System ◽  
Dependent Manner ◽  
Amino Acid Depletion

Abstract The fine-tuning of gene expression contributes to both basic science and applications. Here, we develop a novel gene expression technology termed CRITGI (CRISPR/Transposon gene integration). CRITGI uses CRISPR/Cas9 to integrate multiple copies of the plasmid pTy1 into Ty1 loci, budding yeast retrotransposons. The pTy1 plasmid harbors a Ty1 consensus sequence for integration, a gene of interest with its own promoter and a selection marker gene. Interestingly, the expression of the pTy1 gene in Ty1 loci could be induced in synthetic complete amino acid depletion medium, which could activate the selection marker gene on pTy1. The induction or repression of the gene on pTy1 depended on Ty1 transcription. Activation of the selection marker gene on pTy1 triggered Ty1 transcription, which led to induction of the gene on pTy1. The gene on pTy1 was not transcribed with Ty1 mRNA; the transcription required its own promoter. Furthermore, the trimethylation of histone H3 on lysine 4, a landmark of transcriptionally active chromatin, accumulated at the 5′ end of the gene on pTy1 following selection marker gene activation. Thus, CRITGI is a unique gene regulation system to induce the genes on pTy1 in amino acid depletion medium and utilizes Ty1 transcription to create a chromatin environment favorable for the transcription of the genes on pTy1.

Transient GUS gene expression in Brassica napus electroporated microspores

Plant Science ◽  
1993 ◽  
Vol 93 (1-2) ◽  
pp. 177-184 ◽  
Author(s):  
Marie-Françoise Jardinaud ◽  
André Souvré ◽  
Gilbert Alibert
Keyword(s):  
Gene Expression ◽  
Brassica Napus ◽  
Gus Gene ◽  
Gus Gene Expression

scholarly journals STUDI PERBANDINGAN METODE TRANSFORMASI DNA MENGGUNAKAN VEKTOR Agrobacterium tumefaciens PADA TANAMAN TEBU (Sacharum hybrid)

2007 ◽  
Vol 13 (1) ◽  
pp. 39-44
Author(s):  
Sri Setyati ◽  
Purnama Oktaviandari ◽  
Muhammad Hazmi ◽  
Bambang Sugiharto
Keyword(s):  
Gene Expression ◽  
Embryogenic Callus ◽  
Multiple Shoots ◽  
Suspension Cultures ◽  
Gus Gene ◽  
In Vitro Plants ◽  
Histochemical Observation ◽  
Selection Medium ◽  
Gus Gene Expression

In order to compare transient expression of gus gene driven by CaMV 35S and rice ubiquitin RUBQ2 promoters, a DNA transformation was conducted using embryogenic callus and suspension cultures of sugarcane. The transient gus expression was observed by histochemical staining method. The histochemical observation of GUS activity after co-cultivation showed that RUBQ2 promoter produced high level of clear blue spots both in embryogenic callus and suspension cultures, while the CaMV35S promoter was not detected. The suspension cultures slightly increased transient gus gene expression compared to embryogenic callus. However, the histochemical analysis of regenerated putative transformant plants after 5 successive cycles on the selection medium showed no blue spots of gus gene expression. PCR amplification of DNA for CaMV35 or nptII in putative transformant plants confirmed that there was no integration of the transformed gene in the genome DNA. The results suggested a possibility of somaclonal variation with callus propagation, thus did not produce transformed plants. To avoid the somaclonal variation, the transformation was conducted using in vitro plants and multiple shoots without intervening callus phase. Histochemical observation of infected materials after co-cultivation showed that almost all of the infected materials partially exhibited blue color in the basal region. In case of in vitro plants, they rapidly grow and multiplied in the selection medium, thus the method provided an excellent system for the transformation in sugarcane. The results suggest that in vitro plants as well as multiple shoots need further investigation to be used as target tissues for Agrobacteriummediated transformation in sugarcane.

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