Limitations to the amplification and stability of human tissue-type plasminogen activator expression by chinese hamster ovary cells

Chinese  hamster ovary cell production of recombinant tissue‐type plasminogen activator  (t‐PA) was increased by amplification of cotransfected dihydrofolate reeducates cDNA using stepwise adaptation to increasing methotrexate (MTX) concentrations. The highest producing clones were isolated at 5 μM MTX and yielded 26,000 U/106 cells/day t‐PA (43 μg/106 cells/day). Above 25 μM MTX, cell specific t‐PA production rates became increasingly variable and the cDNA copynumbers decreased. No apparent correlation between the cell specific t‐PA production rate and the growth rate was observed upon sub cloning of the amplified cells. When MTX selection was removed, the t‐PA production rate decreased up to tenfold within 40 days; this was accompanied by an up to 60% drop in cDNA copynumber. Subclones isolated after 108 days of culture in the absence of MTX were, on average, sixfold more stable than their parental cells. In culture without MTX, the maximum stable t‐PA production rate obtained (over 250 days) was 7000 ± 750 U/106cells/day (∼12 μg/106 cells/day), approximately threefold lower than the maximum unstable levels of production reached under selective pressure. Taken together, these results define a wide range of the highest t‐PA expression rates obtained under MTX selection, for which stable expression without selection has not been reported

when you breathe in, or inhale, your diaphragm contracts (tightens) and moves downward. this increases the space in your chest cavity, into which your lungs expand. the intercostal muscles between your ribs also enlarge the chest cavity. they contract to pull your rib cage both upward and outward when you inhale.

a researcher conducts an experiment on the secretion of a particular hormone in mice. scientists inject mice with a substance that stimulates the production of the hormone. the scientists then test the levels of hormones produced by the mice. the tool used to measure the hormones consistently detects the levels at 10 points lower than the actual hormone levels in the mice. this tool makes measurements, but the measurements aren't .

explanation:

no u