scholarly journals Development of endogenous beta-galactosidase and autofluorescence in rat brain microvessels: implications for cell tracking and gene transfer studies.

1994 ◽  
Vol 42 (7) ◽  
pp. 953-956 ◽  
Author(s):  
B Lal ◽  
M A Cahan ◽  
P O Couraud ◽  
G W Goldstein ◽  
J Laterra
Keyword(s):  
Rat Brain ◽  
Reporter Gene ◽  
Cell Tracking ◽  
Fluorescent Dyes ◽  
Tumor Biology ◽  
Lacz Reporter ◽  
Lacz Reporter Gene ◽  
Specific Staining ◽  
Adult Rat ◽  
Beta Galactosidase

Cell transplantation is commonly used in studies of CNS development, tumor biology, and gene therapy. Fluorescent dyes and the E. coli lacZ reporter gene are used to identify transplanted cells in host tissues. The usefulness of these methods depends on host autofluorescence and beta-galactosidase (beta-Gal) activity. Our interest in the CNS vasculature led us to examine vascular autofluorescence and beta-Gal activity in postnatal and adult rat brains. Brains were perfusion-fixed (3.7% paraformaldehyde), cryoprotected, and cryostat-sectioned (12 microns). Autofluorescent vessel profiles were quantitated in sections using rhodamine filter sets and beta-Gal-positive vessels were quantitated under bright-field after incubation of sections with X-Gal chromogenic substrate for 1-18 hr at 37 degrees C. Multifocal vessel autofluorescence appeared in postnatal Day (PND) 18 Lewis rats (0.6 +/- 0.4 vessels/field) and increased tenfold in adults (6.8 +/- 0.3/field). The numbers of beta-Gal-positive vessels in PND 18 and adult sections incubated with X-Gal for 18 hr were 21.1 +/- 1.7 and 119 +/- 17, respectively. Host beta-Gal staining was similar to that produced by implanted endothelial cells expressing the bacterial lacZ reporter gene. Reducing incubation times in X-Gal to less than 4 hr eliminated endogenous staining and retained lacZ-specific staining. The presence of vascular autofluorescence and endogenous beta-Gal activity must be considered when either fluorescence- or lacZ-dependent cell markers are used in rat brain.

scholarly journals The suppressor of Hairy-wing protein regulates the tissue-specific expression of the Drosophila gypsy retrotransposon.

Genetics ◽  
1995 ◽  
Vol 139 (1) ◽  
pp. 215-228 ◽  
Author(s):  
P A Smith ◽  
V G Corces
Keyword(s):  
Reporter Gene ◽  
Leucine Zipper ◽  
Fat Body ◽  
Lacz Reporter ◽  
Lacz Expression ◽  
Lacz Reporter Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Gypsy Retrotransposon ◽  
Beta Galactosidase

Abstract The gypsy retrotransposon of Drosophila melanogaster causes mutations that show temporal and tissue-specific phenotypes. These mutant phenotypes can be reversed by mutations in su(Hw), a gene that also regulates the transcription of the gypsy element. Gypsy encodes a full-length 7.0-kb RNA that is expressed in the salivary gland precursors and fat body of the embryo, imaginal discs and fat body of larvae, and fat body and ovaries of adult females. The su(Hw)-binding region inserted upstream of the promoter of a lacZ reporter gene can induce beta-galactosidase expression in a subset of the embryonic and larval tissues where gypsy is normally transcribed. This expression is dependent on the presence of a functional su(Hw) product, suggesting that this protein is a positive activator of gypsy transcription. Flies transformed with a construct in which the 5' LTR and leader sequences of gypsy are fused to lacZ show beta-galactosidase expression in all tissues where gypsy is normally expressed, indicating that sequences other than the su(Hw)-binding site are required for proper spatial and temporal expression of gypsy. Mutations in the zinc fingers of su(Hw) affect its ability to bind DNA and to induce transcription of the lacZ reporter gene. Two other structural domains of su(Hw) also play an important role in transcriptional regulation of gypsy. Deletion of the amino-terminal acidic domain results in the loss of lacZ expression in larval fat body and adult ovaries, whereas mutations in the leucine zipper region result in an increase of lacZ expression in larval fat body and a decrease in adult ovaries. These effects might be the result of interactions of su(Hw) with activator and repressor proteins through the acidic and leucine zipper domains to produce the final pattern of tissue-specific expression of gypsy.

Promoter Assessment in Hansenula polymorpha Using a lacZ Reporter Gene

2003 ◽  
pp. 101-115
Author(s):  
Manfred Suckow ◽  
Martina Kutzner ◽  
Carsten Amuel ◽  
Cornelis P. Hollenberg ◽  
Gerd Gellissen
Keyword(s):  
Reporter Gene ◽  
Hansenula Polymorpha ◽  
Lacz Reporter ◽  
Lacz Reporter Gene

Nucleosomal location of the STE6 TATA box and Mat alpha 2p-mediated repression

1994 ◽  
Vol 14 (6) ◽  
pp. 4002-4010
Author(s):  
H G Patterton ◽  
R T Simpson
Keyword(s):  
Reporter Gene ◽  
Tata Box ◽  
Chromatin Domain ◽  
Specific Gene ◽  
Multicopy Plasmid ◽  
Alpha Cells ◽  
Lacz Reporter ◽  
Lacz Reporter Gene ◽  
Histone Octamer ◽  
Repression Domain

It has been proposed that yeast MATa cell-specific genes are repressed in MAT alpha cells by the Mat alpha 2p repressor-directed placement of a nucleosome in a position that incorporates the TATA box of the MATa-specific gene close to the nucleosomal pseudodyad. In this study, we address this proposal directly with a series of plasmids designed to place the MATa-specific STE6 TATA box at different locations in a nucleosome and in the internucleosomal linker. These plasmids contain different lengths of synthetic random DNA between the Mat alpha 2p operator and the TATA box of the STE6 promoter, which is located upstream of a lacZ reporter gene in a multicopy plasmid. We show that in MAT alpha cells, a nucleosome is retained in an identical translational frame relative to the Mat alpha 2p operator in all the constructs investigated, irrespective of the sequence of the DNA wrapped onto the histone octamer. This result shows that the nucleosomal organization of the STE6 promoter in MAT alpha cells is not conferred by the sequence of the promoter itself. No expression of the lacZ reporter gene was detectable in MAT alpha cells in any of the constructs, even with the TATA box located in a short internucleosomal linker. These data indicate that repression of MATa-specific genes in MAT alpha cells does not require the precise translational placement of the TATA box close to the nucleosomal pseudodyad; the gene remains repressed when the TATA box is located within the investigated 250-bp region in the organized chromatin domain abutting the Mat alpha 2p operator in MAT alpha cells and may remain repressed with the TATA box located anywhere within this organized repression domain.

Endothelium-specific replacement of the connexin43 coding region by a lacZ reporter gene

genesis ◽  
2000 ◽  
Vol 29 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Martin Theis ◽  
Cor de Wit ◽  
Thorsten M. Schlaeger ◽  
Dominik Eckardt ◽  
Olaf Kr�ger ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Lacz Reporter ◽  
Coding Region ◽  
Lacz Reporter Gene

scholarly journals Moderate Hypermutability of a Transgenic lacZ Reporter Gene in Myc-Dependent Inflammation-Induced Plasma Cell Tumors in Mice

2004 ◽  
Vol 64 (2) ◽  
pp. 530-537 ◽  
Author(s):  
Klaus Felix ◽  
Axel Polack ◽  
Walter Pretsch ◽  
Sharon H. Jackson ◽  
Lionel Feigenbaum ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Plasma Cell ◽  
Lacz Reporter ◽  
Lacz Reporter Gene

A subset of nociceptin/orphanin FQ receptor-expressing neurons in the anterior hypothalamic area, as revealed in mice with lacZ reporter gene

2003 ◽  
Vol 335 (3) ◽  
pp. 217-219 ◽  
Author(s):  
Takeshi Houtani ◽  
Michiko Ikeda ◽  
Masahiko Kase ◽  
Kazuo Sato ◽  
Satoru Sakuma ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Orphanin Fq ◽  
Lacz Reporter ◽  
Lacz Reporter Gene ◽  
Hypothalamic Area
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