Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The introduction of genetic sequences into hematopoietic stem cells (HSC) has allowed study of HSC proliferation in vivo by proviral-sequence molecular analysis in the DNA of progeny. Analysis of HSC proliferation could be enhanced by development of a retroviral vector that encodes a reporter gene that allows sensitive detection of transduced cells. We developed a recombinant retrovirus vector encoding the reporter gene lacZ under the transcriptional control of the myeloproliferative sarcoma virus long-terminal repeat (LTR). Bone marrow cells from C3H mice were co-cultured on retrovirus producer cell lines and cultured for growth of colony-forming unit granulocyte/macrophage (CFU-GM) and high proliferative potential colony-forming cells (HPP-CFC) in semisolid media or were transplanted into irradiated recipients. In other experiments, recombinant retrovirus was injected in vivo into the liver of developing fetal rat pups, and circulating hematopoietic cells of the postnatal rats were analyzed for evidence of proviral integration and expression of beta-galactosidase. Expression of lacZ was detected in both CFU-GM and HPP-CFC that were cultured immediately following in vitro infection of mouse bone marrow. Beta-galactosidase activity from the retrovirus was also detected in both marrow cells isolated from reconstituted mice 22 weeks following transplantation as well as in blood cells of postnatal rats transduced in utero with the recombinant retrovirus. This strategy may be especially useful for characterizing proliferation of transduced populations of hematopoietic cells and in the development of protocols for somatic gene therapy.
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PMID:Myeloproliferative sarcoma virus directed expression of beta-galactosidase following retroviral transduction of murine hematopoietic cells. 760 Dec 55

Human bone marrow mononuclear cells (BMMNCs) and enriched CD34 positive (CD34+) cells were transduced with adenovirus vectors encoding Escherichia coli beta-galactosidase gene. Tranductions were carried out by 24-hour coincubation with adenovirus vectors at different multiplicities of infections (moi). Efficacy of gene transfer into BM cells and expression of the gene product (ie, beta-galactosidase) were studied using X-Gal histochemical staining and flow cytometric analysis. X-Gal staining demonstrated that the percentage of positive cells at mois of 5 to 500 was 3.4% to 34.5% for BMMNCs and 6.0% to 20.0% for enriched CD34+ cells. Similar results (1.5% to 35.7% for BMMNCs and 5.4% to 24.2% for enriched CD34+ cells) were obtained with flow cytometric analysis using fluorescein di-beta-D-galactopyranoside (FDG). Multicolor flow cytometry analysis, which included FDG, demonstrated that BM progenitors (CD34+ or CD34+CD38-), T cells (CD2+), B cells (CD19+), natural killer cells (CD56+), granulocytes, and monocytes all expressed the adenovirus transgene. To ascertain the effects of adenovirus vectors on normal BM progenitors, the numbers of colony forming unit-granulocyte/macrophage (CFU-GM), burst-forming unit-erythrocyte (BFU-E), and high-proliferative potential-colony-forming cells (HPP-CFC) after 24-hour coincubation with adenovirus vectors were determined. When BMMNCs or enriched CD34+ cells were incubated with adenovirus vectors at mois of 5 and 50, no significant differences in the numbers of CFU-GM, BFU-E, and HPP-CFC were observed compared with the uninfected control cells. However, the numbers of CFU-GM were significantly (P < .01) decreased when BMMNCs or enriched CD34+ cells were incubated with adenovirus vectors at a moi of 500, compared with the uninfected control cells. The adenovirus infected cells, purified by cell sorting for FDG expression, were capable of growing in culture and gave rise to various colonies (ie, CFU-GM, BFU-E, and HPP-CFC). These data indicate that recombinant adenovirus vectors can be used to transfer genes to human BM hematopoietic cells with expression of the exogenous gene at a high transduction efficiency.
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PMID:Gene transfer into human bone marrow hematopoietic cells mediated by adenovirus vectors. 919 98