Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:2.4.2.8 (
hypoxanthine-guanine phosphoribosyltransferase
)
2,527
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Valid housekeeping genes (HKG) are a prerequisite for accurate gene quantification. We performed real-time reverse transcription-polymerase chain reaction to investigate the gene expression of five commonly used HKGs (beta-actin, glyceraldehyde-3-phosphate dehydrogenase [GAPDH],
ubiquitin C
[UBC], hypoxanthine phosphoribosyl-transferase [
HPRT
], and cyclophilin A [CYPa]) and antioxidant enzymes in the liver of young and old male Fischer rats. A wide variation in HKG expression existed during the aging process, and
HPRT
was identified as the most stable HKG in rat liver aging. When Cu/Zn-superoxide dismutase gene expression was normalized to
HPRT
, there was no detectable difference between young and old rats; however, a significant difference was seen when it was normalized to UBC. The variation of UBC caused the misinterpretation of Cu/Zn-superoxide dismutase expression. Catalase expression was significantly decreased, whereas glutathione peroxidase expression was not altered with age. We demonstrated that
HPRT
was an appropriate HKG, validation of HKGs was vital for accurate quantification, and decreased catalase expression might be involved in the decline of antioxidant defenses during rat liver aging.
...
PMID:Identification of valid housekeeping genes and antioxidant enzyme gene expression change in the aging rat liver. 1645 91
Progress in our understanding of the molecular cellular basis of immune function depends on our ability to track and image individual immune cells in vivo. To this end, the development of mouse models over-expressing various fluorescent proteins would represent an important experimental tool. In this report, we describe the generation and characterization of pUbi-mRFP-1 transgenic mice, in which the monomeric form of red fluorescent protein is ubiquitously expressed in various lymphoid and non-lymphoid tissues. Our newly generated pUbi-mRFP-1 mice are unique among previously reported mice transgenic for red fluorescent proteins because a single-copy of the mRFP-1 transgene driven by human
ubiquitin C
promoter has been integrated by homologous recombination into the mouse
hypoxanthine-guanine phosphoribosyltransferase
(
HPRT
) locus. We show that the distinct and uniform levels of mRFP-1 expression allow easy identification of transferred hematopoietic cells by FACS analysis or confocal microscopy, even when the transferred population represents a very small proportion in the target organ. Also, even in long-term experiments, we have seen no evidence of rejection of transferred pUbi-mRFP-1 lymphocytes. Due to its far-red spectrum, mRFP-1 is an ideal partner for dual imaging with green fluorescent proteins. We observed a good visual separation between donor lymphocytes derived from either mRFP-1 or eGFP transgenic mice in recipient animals. Our study suggests that the new pUbi-mRFP-1 transgenic mouse strain offers new opportunities for studying cellular interactions and migratory patterns of cells, especially for dual imaging of different cell types. In summary, our results demonstrate that a controlled strategy of transgenesis provides an effective means of ubiquitously expressing fluorescent proteins in vivo.
...
PMID:Ubiquitous expression of mRFP-1 in vivo by site-directed transgenesis. 1707 85
Comprehensive analyses of gene expression have been carried out by the development of microarrays and deep sequencers. However, it is difficult to obtain comprehensive information on gene expression from a small amount of ribonucleic acid (RNA). Therefore, we investigated the reproducibility and application of T7 RNA polymerase-mediated transcription, adaptor ligation and polymerase chain reaction (PCR) amplification, followed by T7 transcription (TALPAT), an efficient method for amplifying poly (A)-positive RNA, such as messenger RNA (mRNA). When amplified complementary RNA (cRNA) was electrophoresed, a large number of amplified cRNA was detected in the size of 0.2-0.5 kb. This indicates that the region up to 0.2-0.5 kb from the 3' end of the original mRNA was amplified by the TALPAT method. Seven housekeeping genes, glyceraldehyde-3-phosphate dehydrogenase (
GAPDH
), hydroxymethylbilane synthase (
HMBS
),
hypoxanthine phosphoribosyltransferase
(
HPRT1
), ribosomal protein L13a (
RPL13A
), succinate dehydrogenase complex (
SDHA
), TATA box-binding protein (
TBP
) and
ubiquitin C
(
UBC
), showed high reproducibility (square of the correlation coefficient, R
2
=0.9954), according to scatter plots of Ct values obtained in the real-time PCR analysis of amplified cRNA. In addition, relative expression ratios of amplified cRNA of the seven housekeeping genes were approximately equal to the ratio of the original RNA solution. Furthermore, cRNA was amplified from 20 pg total RNA. In the present study, we confirmed the characteristics of mRNA amplification using the TALPAT method. This method may be applicable to mRNA and poly (A)-positive non-coding RNA amplification, using a small amount of RNA from single, laser-captured and sorted cells, as well as exosomes from serum, urine and body fluids.
...
PMID:An efficient method for high-fidelity messenger RNA amplification from a small amount of total RNA. 2464 3
