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ConstructionofRecombinantAdenovirusofhAIF-1GeneandItsEffectonProliferationandMigrationofA549Cells(PDF)

《南京师大学报(自然科学版)》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2015年03期
Page:
64-
Research Field:
生命科学
Publishing date:

Info

Title:
ConstructionofRecombinantAdenovirusofhAIF-1GeneandItsEffectonProliferationandMigrationofA549Cells
Author(s):
LuoLiyunBaiMinZhangJianCaoLingsenCaoXiangrong
SchoolofLifeSciences,NanjingNormalUniversity,JiangsuProvinceKeyLaboratoryforMolecularandMedicalBiotechnology,Nanjing210023,China
Keywords:
hAIF-1adenovirusproliferationmigration
PACS:
Q28
DOI:
-
Abstract:
hAIF-1isaninflammatoryfactor.ThestudywassupposedtoexploretheeffectofhAIF-1ontheproliferationandmigrationofA549cellsbythepreparationoftherecombinantadenoviruswithhAIF-1gene.ThehAIF-1genewasclonedbyPCRandsubclonedintotheshuttlevectorpDC315-EGFP.TheshuttleplasmidandadenovirusgenomicplasmidpBHGloxdeltaE13Crewereco-transfectedinto293cellsforpackagingrecombinantadenoviruswiththehelpoftheadenoviruspackagingsystemAdMax.ThetiteroftheviruswasmeasuredbyTCID50.TheexpressionofthetargetgenewasidentifiedbyRT-PCRandWesternBlot,andtheeffectofthetargetgeneontheproliferationandmigrationofA549cellswasexaminedbyMTT,woundhealingandTranswellassay.TheresultsshowedthattheAdDC315-EGFP-hAIF-1wassuccessfullyconstructedandthetiteroftheviruswas2.5×108pfu/mL;AnalysisofRT-PCRandWesternBlotindicatedthatA549cellscouldoverexpresshAIF-1afterinfectedwiththerecombinantadenovirus;TheMTTassayshowedthathAIF-1couldincreasetheproliferativerateofA549cells;ThewoundhealingandTranswellassayprovedthathAIF-1couldpromotethemigrationofA549cells.

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Last Update: 2015-09-30