Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique-中国Betway体育网页登陆

摘要 Themethodoflasercapturemicrodissection(LCM)combinedwithsuppressivesubtractivehybridization(SSH)wasdevelopedtoisolatespecificgermcellsfromhumantestissectionsandtoidentifythegenesexpressedduringdifferentiationanddevelopment.Inthepresentstudy,over10,000primaryspermatocytesandroundspermatidcellsweresuccessfullyisolatedbyLCM.UsingthecDNAsfromprimaryspermatocytesandroundspermatids,SSHcDNAslibraryofprimaryspermatocyte-specificwasconstructed.TheaverageinsertsizeofthecDNAisolatedfrom75randomlypickedwhitecloneswas500bp,rangingfrom250bpto1.7kb.Usingthedot-blotmethod,atotalof421cloneswereexamined,resultingintheidentificationof390positiveclonesemittingstrongsignals.PartialsequenceofcDNAspreparedfromeachclonewasdeterminedwithanoverallsuccessrateof84.4%.GenesencodingcytochromecoxidaseⅡandtherescuefactor-humaninweremostfrequentlyexpressedinprimaryspermatocytes,suggestingtheirrolesinvolvedinmeiosis.

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Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique

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