克隆测序法鉴定SCN5A基因多态性与克山病相关性研究的开题报告.docx

克隆测序法鉴定SCN5A基因多态性与克山病相关性研究的开题报告

Title:IdentificationofSCN5AgenepolymorphismsusingcloningsequencingmethodandtheircorrelationwithKashin-Beckdisease

Introduction:

Kashin-Beckdisease(KBD)isachronicjointdiseasewhichmainlyaffectschildrenandyoungadults.ItishighlyassociatedwithseleniumdeficiencyandisprevalentincertainareasofChina.TheetiologyofKBDisnotclear,butithasbeensuggestedthatgeneticfactorsplayaroleinitsdevelopment.SCN5Agene,whichencodesthesodiumchannelNav1.5,hasbeenlinkedwithvariouscardiovascularconditions,anditishypothesizedthatitmayalsocontributetothepathogenesisofKBD.

Objective:

TheobjectiveofthisstudyistoidentifyandanalyzethepolymorphismsofSCN5AgeneusingcloningsequencingmethodandinvestigatetheircorrelationwithKBD.

Methods:

PeripheralbloodsampleswillbecollectedfromKBDpatientsandhealthycontrols.GenomicDNAwillbeextractedandPCRprimerswillbedesignedtoamplifytheSCN5Agene.ThePCRproductswillthenbeclonedintothepUC19vectorandtransformedintocompetentE.colicells.Thepositivecolonieswillbeselectedandsentforsequencing.Theobtainedsequenceswillbecomparedwiththereferencesequencetoidentifypolymorphisms.ThegenotypesandallelefrequenciesofeachpolymorphismwillbecalculatedandcomparedbetweenKBDpatientsandcontrols.

Expectedresults:

ItisexpectedthatseveralpolymorphismsofSCN5Agenewillbeidentifiedinthisstudy.ThegenotypeandallelefrequenciesofeachpolymorphismwillbeanalyzedtoinvestigatetheircorrelationwithKBD.ItishypothesizedthatcertainpolymorphismsofSCN5AgenemaybeassociatedwithKBD,whichmayprovideinsightsintothepathogenesisofthisdisease.

Conclusion:

ThisstudyaimstoidentifyandanalyzethepolymorphismsofSCN5AgeneusingcloningsequencingmethodandinvestigatetheircorrelationwithKBD.Thefindingsofthisstudymayprovidenewins