Posts Tagged ‘expression’
Last Updated on Friday, 27 August 2010 12:34 Written by Administrator Friday, 27 August 2010 12:34
Our vision at Aldevron is to be a world-class global contract services organization. To achieve this, we need to understand what our clients need and deliver what they want. One of our services is expression screening in insect cells. We offer a range of options to suite the clients needs. A typical screening project includes multiple variants, such as amino acid changes or truncations, of a single gene. We are able to screen multiple variants and culture conditions in parallel. There are a couple of options for expression screening in insect cells. The first option is to screen for expression via baculovirus infection. We can either start with a recombinant transfer vector supplied by the client or we can clone the gene of interest into a baculovirus transfer vector. Next, we generate a high-titer baculovirus stock and determine the viral titer. We then infect 10ml Sf21 cultures for recombinant protein expression. The second option is to screen for expression via direct transfection. This allows us to go from plasmid to protein in as little as 48 hours. We start by cloning the gene of interest into an expression vector designed to enable robust expression in insect cells without the need to produce recombinant baculovirus. Next we transfect 10ml Sf9 cultures with the recombinant plasmids. 48-72 hours post-transfection, we harvest the cells. Because generating a recombinant baculovirus can require anywhere between 2-4 weeks of time, the direct transfection method offers a …
Tags: Aldevron, Cells, expression, Insect, Protein | Posted under Western Blot Videos | No Comments
Last Updated on Sunday, 18 July 2010 07:45 Written by Administrator Sunday, 18 July 2010 07:45
DNA polymerase @b mRNA and protein expression in Xiphophorus fish [An article from: Comparative Biochemistry and Physiology, Part C]
This digital document is a journal article from Comparative Biochemistry and Physiology, Part C, published by Elsevier in 2004. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
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Herein we report Xiphophorus DNA polymerase @b (XiphPol@b) mRNA and protein expression levels in brain, liver, gill, and testes tissues from Xiphophorus maculatus, Xiphophorus helleri, and Xiphophorus couchianus parental line fish and two different tumor-bearing Xiphophorus interspecies hybrids. Polymerase @b protein levels in the Xiphophorus tissues were measured by Western blot, and mRNA was measured with a quantitative real time RT-PCR method which employed cRNA construction to produce accurate calibration curves. We found significant differences in both mRNA and protein levels between the tumor-bearing hybrid animals and the three parental species. However, there were no significant differences in either mRNA levels or protein expression observed between the parental species. Thus, interspecies hybridization results in dysregulation of Pol@b expression and this may manifest a modulation in DNA repair capability and susceptibility to latent tumorigenesis.
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Tags: article, Biochemistry, Comparative, expression, fish, from, mRNA, Part, Physiology, polymerase, Protein, Xiphophorus | Posted under Western Blot Books | No Comments
Last Updated on Sunday, 18 July 2010 07:45 Written by Administrator Sunday, 18 July 2010 07:45
MAP2 and synaptophysin protein expression following motor learning suggests dynamic regulation and distinct alterations coinciding with synaptogenesis … from: Neurobiology of Learning and Memory]
This digital document is a journal article from Neurobiology of Learning and Memory, published by Elsevier in 2007. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
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Learning a new motor skill can induce neuronal plasticity in rats. Within motor cortex, learning-induced plasticity includes dendritic reorganization, synaptogenesis, and changes in synapse morphology. Behavioral studies have demonstrated that learning requires protein synthesis. It is likely that some of the proteins synthesized during learning are involved in, or the result of, learning-induced structural plasticity. We predicted the expression of proteins involved in neural plasticity would be altered in a learning dependent fashion. Long-Evans rats were trained on a series of motor tasks that varied in complexity, so that the effects of activity could be teased apart from the effects of learning. The motor cortices were examined for MAP2 and synaptophysin protein using Western blotting and immunohistochemistry. Western blotting revealed that expression of MAP2 was not detectably influenced by learning, whereas synaptophysin expression increased on day 1, 3, and 5 of complex motor skill learning. Expression of MAP2 does not seem to indicate difficulty of task or duration of training time, whereas increases in synaptophysin expression, which appear diffusely across the cortex, seem to be correlated with the first 5 days of motor skill learning. Similar findings with GAP-43 suggest the change in synaptophysin may coincide with synapse formation. Immunohistochemistry did not reveal any localized changes in protein expression. These data indicate a difference in learning-induced expression in the mammalian brain compared to reports in the literature, which have often focused on stimulation to induce alterations in protein expression.
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Tags: alterations, coinciding, distinct, dynamic, expression, following, from, learning, MAP2, Memory, motor, Neurobiology, Protein, regulation, suggests, synaptogenesis, synaptophysin | Posted under Western Blot Books | No Comments
Last Updated on Sunday, 18 July 2010 07:44 Written by Administrator Sunday, 18 July 2010 07:44
Bacterial infection and tissue-specific Hsp72, -73 and -90 expression in western painted turtles [An article from: Comparative Biochemistry and Physiology, Part C]
This digital document is a journal article from Comparative Biochemistry and Physiology, Part C, published by Elsevier in 2004. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
Description:
Heat shock proteins (Hsps) are molecular chaperones that assist intracellular folding, assembly and translocation of proteins in prokaryotic and eukaryotic cells. A variety of stresses including hyperthermia, radiation, heavy metals, ischemia, anoxia and reoxygenation have been shown to increase the expression of Hsps. Likewise, bacterial infection represents a stress for the host cell. In this study, expression of the constitutive (Hsp73) and inducible (Hsp72) isoforms of Hsp70 and Hsp90 was monitored in brain, heart, liver and skeletal muscle from the western painted turtle Chrysemys picta bellii diagnosed with Septicemic Cutaneous Ulcerative Dermatitis (SCUD). This disease is caused by a gram-negative bacterium probably belonging to the Citrobacter spp. The expression of Hsp73 increased 1.8-fold in brain and liver, 2.2-fold in heart but did not change in skeletal muscle; Hsp72 expression increased 5.5-fold in brain and 3-fold in liver but did not change in heart or skeletal muscle; Hsp90 expression increased 9-fold in brain, 2.7-fold in heart and 2.4-fold in skeletal muscle but did not change in liver. These results suggest a tissue-specific Hsp response during bacterial infection and a role for Hsps in immunopathological events in reptiles.
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Tags: article, Bacterial, Biochemistry, Comparative, expression, from, Hsp72, infection, painted, Part, Physiology, tissuespecific, turtles, Western | Posted under Western Blot Books | No Comments