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Expression, purification and cellular uptake of a S100A4 protein. an agent promoting metastasis

Автор:   stian s.
мягкая обложка
152 страниц
2009 год
Издательство:   Книга по Требованию
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2382 руб
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Описание:A recombinant S100A4 protein was produced in Pichia pastoris (Pichia). Similar to a mouse rS100A4 produced in bacterial cells, the Pichia produced protein was also found internalized into the HeLa cells. The protein eluted from the hydrophobic column was labelled with the fluorescent dye (Cy3) and used for internalization studies in the human cell line HeLa. The expression vector contained parts of the AOX1 promoter and also a secretion signal, making it possible to purify the protein from the yeast media after methanol induction. The biological significance of these findings is at present not elucidated and will be further studied. Attempts were made to purify the protein from the media both by hydrophobic interaction chromatography (HIC) and by anionic ion exchange chromatography (IEC). The binding capacity to the hydrophobic material was rather low, but the eluted protein did however show an acceptable purity for utilization in downstream functional assays.


Orthomol Immun Junior Сила иммунитета на страже Вашего ребенка Orthomol Immun Junior Сила иммунитета на страже Вашего ребенка

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A short history of nearly everything

As Morgan noted at the time, there was no consensus БЂњas to what the genes are-whether they are real or purely fictitious.БЂ« It may seem surprising that scientists could struggle to accept the physical reality of something so fundamental to cellular activity, but as Wallace, King, and Sanders point out in Biology: The Science of Life (that rarest thing: a readable college text), we are in much the same position today with mental processes such as thought and memory. We know that we have them, of course, but we donБЂ™t know what, if any, physical form they take. So it was for the longest time with genes. The idea that you could pluck one from your body and take it away for study was as absurd to many of MorganБЂ™s peers as the idea that scientists today might capture a stray thought and examine it under a microscope. What was certainly true was that something associated with chromosomes was directing cell replication. Finally, in 1944, after fifteen years of effort, a team at the Rockefeller Institute in Manhattan, led by a brilliant but diffident Canadian named Oswald Avery, succeeded with an exceedingly tricky experiment in which an innocuous strain of bacteria was made permanently infectious by crossing it with alien DNA, proving that DNA was far more than a passive molecule and almost certainly was the active agent in heredity ...»

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