微生物学微生物学 (49).docVIP

00:14

Hieveryone,inthissectionwewilltalkaboutgeneengineering.Afterwatchingthisvideo,youshouldbeabletotelltheworkflowofgeneticengineering.

00:32

Geneengineering,alsocalledgeneticengineering.Nowadays,itisawidelyusedtechnologyinmodernbiology.WeknowthatRecombinantDNAisDNAwithanewnucleotidesequence.SuchDNAisformedbyjoiningfragmentsfromtwoormoredi?erentsources.Inthissection,wewillfocusontheworkflowofgeneticengineering.

1:13

Thebasicstepsofgeneticengineeringcanbebrieflydescribedasshowninthisfigure.

Firstly,isolatingDNAtobecloned;Secondly,usearestrictionenzymeorPCRtogeneratefragmentsofDNA;andthirdly,generatingarecombinantmoleculebyinsertingDNAfragmentsintoavector;andfourthly,introducingrecombinantmoleculeintoanewhost;Finally,screeningthesuccessfullytransformedhost,generallytheywillexhibitanewtraitduetotheexpressionofthetransformedgene.

2:17

Toobtaintargetgeneofinterestisthefirstkeystepforgeneticengineering.Thetargetgenecanbeextractedfrommicrobe,animalorplant.ItcanalsobesynthesizedofcDNAfrommRNAviareversetranscriptase,orchemicallysynthesized.

2:49

Thenextstepisselectingthesuitablegeneticvectorfortransformation;thereareseveralrequirementswhilechoosingthevector:firstly,itshouldbeareplicatorwithlowmolecularweight,itcanreplicate;secondly,itcanmultiplyinreceptorcellstoalargenumber;thirdly,itusuallyhaveonlyonerestrictiveendoenzymesitesothatthetargetgenecanbeintegratedintothevectorDNA;fourthly,theremustbeageneticmarkerforefficientscreen.Forexample,somevectorshaveantibiotics-resistancesitesothattheycansurviveaftergrowingonmediumwithantibiotics.

4:06

Thenextstepisintegratingthetargetgenewiththegeneticvector,DNAsequenceofbothtargetgeneandgeneticvectorcanbecatalyzedbyEndonucleases