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Virofection: a new procedure to achieve stable expression of genes transferred into early embryos

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Figure

Fig. 2. Structureof theposedhelperandvectorplasmids.Cisroravis com-of an assemblyof the non-codmgsequencesof a mUflne retrovirus(hatchedboxes)unrelatedto avianleukosisVlfuses,andthecodingsequencesof rhe Rous associatedvIrus type-I.The defectiveretrovirusgenomeL210 derivedfrom Rous sarcomaVlfUS is shownbelow.pBlagy3wasconstructedby replacingmostof thecodmgsequencesof Rousassociatedvirustype-2by a lacZ-NeoRfusiongene.DottedImes corre-spond to the homologiesfound betweenCisroravandthe vecrorgenomes.in the gag codmgsequenceand at the end of the env sequence.
Fig. 3. Virofectionof theplasmid)lacZgenein CEF in vitro.(AI T/mecourse'whirerriangles:cc-rransfecrionof a 60/40%mi;...rure of Osrorav(helperand pLZl 0 (vectorplasmidcarryingthe lacZ gene).Black triangles:transfect/onof pLZ10alone.The frequencyof CEF displayinglacZ e>o.'pres-sion in siru (lac+ cells) /s plottedon a logarithmicscale.(8) Influenceof therelativeratio of pLZ10and Ostoravon rhe frequencyof lac+ cells by day 9.
Fig. 5. Structureof the lacZ gene integratedin CEFaftervirofectionisreminiscentof a retrovirus-mediatedintegrationprocess.CEF DNAt'!l.tracted 76 daysafterInvirroVlfofectlon
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