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Potent transposase-dependent integration of genetic cargo carried by the hybrid HIV-1/SB vector (up to 160-fold above background) is reported in human cell lines as well as in primary human fibroblasts and keratinocytes.
In good agreement, durable increases in the proteoglycan and type-II collagen contents were noted in dynamic versus static culture conditions at any time point examined, with or without vector (up to 1.8- and 1.7-fold difference for the proteoglycan and type-II collagen contents, respectively, P ≤ 0.001) (Tables 1 and 2).
In order to identify HAsik1 or p57 overexpressing ES cells, two weeks after selection with 1.2 µg/mL puromycin, the resistant colonies were screened by RT-PCR using the primers sik1UP and sik1LW for HAsik1 clones and with vector up and p57low for the p57 clones.
The amount of total DNA was normalized with empty pcDNA3 vector (up to 100 ng).
COS7 cells were transfected in 24-well plates with 20 ng of HA-tagged mouse RHBDL2-KDEL (RHBDL2) plasmid, 50 ng of FLAG-tagged Gurken substrate construct, and pcDNA3 vector up to a total amount of 200 ng DNA per well using Fugene6 (Roche).
Ad-mIL-22 resulted in the complete protection as compared with control vector up to 20 days after treatment.
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This is typically not a problem in simple projects using recipient vectors up to several thousand bases (kb) long.
In a previous experimental study, we showed that the assay for guinea pig DNA was successful for 100% of the vectors up to 40 hours after bugs had fed.
The assay detects DNA quite soon after feeding, we detected guinea pig DNA in 100% of the vectors up to 40 hours after bugs had fed on guinea pigs under controlled conditions.
Although DNA from the vector turned up in other tissues, the amounts were extremely low.
Unless otherwise noted, we denote w the vector made up by the column stacking of the vectors wk,n.
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