A Method for Obtaining Reporter Gene Activity and Nuclear Extracts Simultaneously from Transiently Transfected Cells

1998 ◽  
Vol 265 (1) ◽  
pp. 185-187 ◽  
Author(s):  
Sanjaya Singh ◽  
Grady F. Saunders
Keyword(s):  
Reporter Gene ◽  
Gene Activity ◽  
Transfected Cells ◽  
Reporter Gene Activity ◽  
Nuclear Extracts

Isotopic Assays for Reporter Gene Activity

2001 ◽  
Vol 63 (1) ◽  
Author(s):  
Robert E. Kingston ◽  
Jen Sheen ◽  
David Moore
Keyword(s):  
Reporter Gene ◽  
Gene Activity ◽  
Reporter Gene Activity

scholarly journals Alternative promoters and repetitive DNA elements define the species-dependent tissue-specific expression of the FMO1 genes of human and mouse

2007 ◽  
Vol 406 (3) ◽  
pp. 491-499 ◽  
Author(s):  
Elizabeth A. Shephard ◽  
Pritpal Chandan ◽  
Milena Stevanovic-Walker ◽  
Mina Edwards ◽  
Ian R. Phillips
Keyword(s):  
Reporter Gene ◽  
Adult Mouse ◽  
Direct Repeat ◽  
Mouse Gene ◽  
Gene Activity ◽  
Upstream Sequence ◽  
Alternative Promoters ◽  
Specific Expression ◽  
Tissue Specific ◽  
Reporter Gene Activity

In humans, expression of the FMO1 (flavin-containing mono-oxygenase 1) gene is silenced postnatally in liver, but not kidney. In adult mouse, however, the gene is active in both tissues. We investigated the basis of this species-dependent tissue-specific transcription of FMO1. Our results indicate the use of three alternative promoters. Transcription of the gene in fetal human and adult mouse liver is exclusively from the P0 promoter, whereas in extra-hepatic tissues of both species, P1 and P2 are active. Reporter gene assays showed that the proximal P0 promoters of human (hFMO1) and mouse (mFmo1) genes are equally effective. However, sequences upstream (−2955 to −506) of the proximal P0 of mFmo1 increased reporter gene activity 3-fold, whereas hFMO1 upstream sequences (−3027 to −541) decreased reporter gene activity by 75%. Replacement of the upstream sequence of human P0 with the upstream sequence of mouse P0 increased activity of the human proximal P0 8-fold. Species-specific repetitive elements are present immediately upstream of the proximal P0 promoters. The human gene contains five LINE (long-interspersed nuclear element)-1-like elements, whereas the mouse gene contains a poly A region, an 80-bp direct repeat, an LTR (long terminal repeat), a SINE (short-interspersed nuclear element) and a poly T tract. The rat and rabbit FMO1 genes, which are expressed in adult liver, lack some (rat) or all (rabbit) of the elements upstream of mouse P0. Thus silencing of FMO1 in adult human liver is due apparently to the presence upstream of the proximal P0 of L1 (LINE-1) elements rather than the absence of retrotransposons similar to those found in the mouse gene.

scholarly journals Human Serum Induces Streptococcal C5a Peptidase Expression

2009 ◽  
Vol 77 (9) ◽  
pp. 3817-3825 ◽  
Author(s):  
Ute Gleich-Theurer ◽  
Simone Aymanns ◽  
Gregor Haas ◽  
Stefanie Mauerer ◽  
Julia Vogt ◽  
...  
Keyword(s):  
Human Serum ◽  
Reporter Gene ◽  
Bovine Serum ◽  
Calf Serum ◽  
Gene Activity ◽  
Pcr Analysis ◽  
Reporter Gene Activity ◽  
Reverse Transcription Pcr ◽  
Bovine Strain ◽  
Human Infections

ABSTRACTStreptococcus agalactiaeis a major pathogen in humans and animals. Virulence factors are often associated with mobile genetic elements, and their expression can be modulated by host factors.S. agalactiaeharbors the genes for C5a peptidase (scpB) and Lmb on a composite transposon structure which is absent in many bovine isolates. To investigate whether these genes participate in the adaptation to human hosts, we determined the influence of human and bovine serum on the promoter activity ofscpBandlmbby using fluorescence-activated cell sorter analysis. Culture in the presence of 1 to 50% human serum resulted in a dose-dependent induction of reporter gene activity forscpBbut notlmb. Reporter gene activity was, however, unchanged following growth in fetal calf serum. Interestingly, a bovine strain did not display any induction ofscpBby either bovine or human serum. Reverse transcription-PCR analysis was used to confirm differential induction ofscpBinS. agalactiaeand showed a similar induction of theStreptococcus pyogenesC5a peptidase genescpAby human but not bovine serum. The specific induction of the streptococcal C5a peptidase by human serum corresponds to the absence ofscpBin many bovineS. agalactiaeisolates and underlines the importance of this virulence factor for human infections.

scholarly journals A simple protocol for preparation of a liposomal vesicle with encapsulated plasmid DNA that mediate high accumulation and reporter gene activity in tumor tissue

2011 ◽  
Vol 1 (1) ◽  
pp. 49-56 ◽  
Author(s):  
Torben Gjetting ◽  
Thomas Lars Andresen ◽  
Camilla Laulund Christensen ◽  
Frederik Cramer ◽  
Thomas Tuxen Poulsen ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Plasmid Dna ◽  
Tumor Tissue ◽  
Gene Activity ◽  
High Accumulation ◽  
Reporter Gene Activity ◽  
Simple Protocol

scholarly journals Transcriptional activation of cathepsin D gene expression by growth factors

2000 ◽  
Vol 24 (2) ◽  
pp. 193-202 ◽  
Author(s):  
F Wang ◽  
R Duan ◽  
J Chirgwin ◽  
SH Safe
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Reporter Gene ◽  
Cathepsin D ◽  
Mapk Pathway ◽  
Gene Promoter ◽  
Gene Activity ◽  
Reporter Gene Activity ◽  
Igf I ◽  
Mcf 7

Insulin-like growth factor-I (IGF-I), transforming growth factor alpha (TGFalpha) and epidermal growth factor (EGF) induced cathepsin D gene expression and reporter gene activity in MCF-7 human breast cancer cells transiently transfected with a construct (pCD1) containing a -2576 to -124 cathepsin D gene promoter insert. In contrast, IGF-I, but not TGFalpha or EGF, induced reporter gene activity in cells cotransfected with wild-type estrogen receptor (ER) expression plasmid and a construct (pCD2) containing estrogen-responsive downstream elements from -208 to -101. Promoter deletion and mutational analysis experiments identified four GC-rich sites and an imperfect palindromic estrogen responsive element required for IGF-I activation of the ER (ligand-independent). Subsequent studies with the mitogen-activated protein kinase (MAPK) inhibitor, PD98059, and a serine(118(-ER mutant confirmed the role of the MAPK pathway for IGF-I activation of the ER in MCF-7 cells. Thus, growth factor activation of ER can mediate transactivation vs ER/Sp1 binding to GC-rich sites and represents a novel pathway for ligand-independent ER action. The divergent pathways for IGF-I and TGFalpha/EGF activation of the ER observed in MCF-7 cells contrast with previous data indicating that pathways for growth factor activation of the ER are dependent on the gene and/or gene promoter and on cell context.

Sign in / Sign up

Export Citation Format

Share Document