Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The HIV-1 Vpu protein stimulates virus production by enhancing the release of viral particles from infected cells. Interestingly, Vpu was also shown to enhance the release of capsids produced by gag gene contructs of other retroviruses that lack a Vpu-like activity. To investigate the effect of Vpu expression on viral particle production in retroviral packaging cell line, we developed the Damp-VpuP cell line in which vpu expression is under the control of the tetracycline-responsive promoter. Retroviral production was measured by dosage of virion-associated reverse transcriptase activity, by capsid protein immuno-detection in cell-free supernatants and by evaluating the transfer of antibiotic resistance to target cells. Induction of the Damp-VpuP cell line caused a 40-fold increase in the titer of infectious virus-like particles when compared with control cell lines. This increase in viral titer was not the result of a clonal effect nor was it a consequence of high selective pressure but rather the effect of a Vpu-mediated enhancement of viral particle production. Similar results using the third generation psi CRIP packaging cell line confirmed these findings. Constitutive expression of vpu caused a 13-fold increase in viral titer in this packaging cell line. These results indicate that the expression of HIV-1 vpu in retroviral packaging cell lines can significantly improve the titers of infectious retroviral particles.
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PMID:Enhancement of retroviral production from packaging cell lines expressing the human immunodeficiency type 1 VPU gene. 933 17

Although recombinant retroviruses are widely used in gene therapy and as gene transfer vehicles for basic biological studies, their titers are very low as compared to other recombinant viral systems, e.g., adenovirus. We investigated the rate-limiting steps in production of LacZ-encoding ecotropic (CRE BAG 2) and amphotropic (Psi-CRIP) retrovirus. We found that ecotropic retrovirus producer cells produced a large number of inactive viral particles because they were severely limited by the amount of mRNA that was packaged into viral capsids. Introduction of the gene for green fluorescence protein (GFP) increased retroviral titers 40-fold, without affecting the viral matrix protein, p30, or the activity of reverse transcriptase. Surprisingly, while transfer of GFP gene increased retrovirus production, beta-gal activity and X-gal titer decreased significantly. Quantitative real-time polymerase chain reaction (PCR) showed that although producer cells synthesized similar amounts of both mRNAs, retroviral supernatants contained significantly lower amount of LacZ mRNA, possibly due to competition between LacZ and GFP mRNAs for encapsidation into virions. In contrast to ecotropic producers, introduction of GFP gene copies into amphotropic producers resulted in a moderate twofold increase in retrovirus production. However, delivery of genes encoding for the viral proteins gp70 and p30 increased virus production by fivefold, suggesting that amphotropic producers may also be limited by synthesis of structural viral proteins. Our data show that in addition to the amount of viral genome or proteins, assembly of viral components into active viral particles may limit production of high titer retroviral preparations.
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PMID:Stoichiometric limitations in assembly of active recombinant retrovirus. 1581 99

Cysteine-rich intestinal protein 1 (CRIP1) is an important transcriptional regulation factor during the tumor development. Although it was largely studied in the human or mouse, no report has provided functional evidence for it in the swine. To date, the real sequence of porcine CRIP1 (poCRIP1) was also still unknown. In this study, clear characteristics for the poCRIP1 were represented. A 552bp poCRIP1 cDNA was obtained from porcine brain tissue using real time reverse transcriptase PCR. The poCRIP1 showed 89% and 93% homologous with human and cattle, respectively. And it also contained one conserved domain, LIM-CRIP domain. Meanwhile, the genomic structure and promoter map was done and several conserved transcriptional regulatory sites were also predicted in this study. The expression pattern of poCRIP1 indicated that poCRIP1 is expressed in mucosal tissue. An infection experiment about the gut was designed to analyze whether or not poCRIP1 was functional in gut immunity, and an interesting result was that poCRIP1 was only activated by an opportunistic pathogen, Enterococcus faecalis FA2-2. It was the first report to identify the full-length sequence of poCRIP1 gene, represent a clear characteristic and immunologic role of CRIP1 in domestic animal until now.
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PMID:CRIP1, a novel immune-related protein, activated by Enterococcus faecalis in porcine gastrointestinal epithelial cells. 2783 62