Korhan ARSLAN, Haydar BAĞIŞ, Munis DÜNDAR

EFFECTS OF SEPERATE OR SIMULTANEOUS INJECTION OF TWO DIFFERENT GENES (ENHANCED GREEN-FLUORESCENCE PROTEIN GENE, HUMAN GAMMA INTERFERON GENE)

hIFN-γ and EGFP gene constructs were separatelyor simultaneously injected to mouse embryos atpronuclear phase. Micro-injection were classifiedin three different experiment groups according toalone or simultaneous transfer of gen constructs. Infirst group co-injection of hIFN-γ+EGFP transgenes were performed. As a result of transfers,totally 39 offsprings were born, In molecularanalysis performed on DNAs obtained from thoseoffsprings, two were EGFP transgenic and one washIFN-γ transgenic. In second group micro-injectionof hIFN-γ trans-gene was performed. As a result oftransfers, totally 21 offsprings were born. Inmolecular analysis performed on DNAs obtainedfrom those offsprings totally two neonates werehIFN-γ transgenic. In third group micro-injection ofEGFP trans-gene was performed. As a result oftransfers, totally five offsprings were born, it wasfound that there were no transgenic offspring.Results presented in present study found thattransgenicity percentage of offsprings born as aresult of simultaneous micro-injection of twodifferent genes was high. If the protocol presentedin this study is applied to transgenic mice recoverystudies, it is possible to recover transgenic micecarrying two different genes separate. Thus, it willbe possible to save money and time

Anahtar Kelimeler:

Transgen, mikroenjeksiyon, EGFP, hIFN-γ, Ko-enjeksiyon

İki Farklı Genin (Güçlendirilmiş Yeşil Floresan Protein Geni, İnsan Gamma İnterferon Geni) Ayrı Ayrı veya Birlikte Mikroenjeksiyonunun Transgenik Fare Eldesi Üzerine Etkilerinin Araştırılması

Sunulan çalışmada hIFN-γ ve EGFP genkonstraktlarının farklı konsantrasyonları birlikte veayrı ayrı pronüklear safhadaki fare embriyolarınamikroenjekte edilmiştir. Mikroenjeksiyon deneyleriaktarılan gen konstraktlarının yalnız ve/veya birlikteaktarımlarına göre üç farklı deney grubu çalışılmıştır.I.Deney Grubunda hIFN-γ ve EGFP genleri birliktemikroenjekte edilmiş, aktarımlar sonucunda toplam 39yavru doğmuştur. Yapılan moleküler analizlerdetoplam üç yavrunun transgenik olduğu belirlenmiştir.Üç transgenik yavrudan iki’si EGFP transgenik, birtanesi ise hIFN-γ transgeniktir. II. Deney Grubundayalnız hIFN-γ geni mikroenjekte edilmiş, aktarımlarsonucunda toplam 21 yavru doğmuştur. Yapılanmoleküler analizlerde toplam iki yavrunun hIFN-γtransgenik olduğu belirlenmiştir. III.Deney Grubundayalnız EGFP geni mikroenjekte edilmiş, aktarımlarsonucunda yavrulardan elde edilen DNA’lar ile yapılan moleküleranalizlerde Sunulan çalışma da elde edilen sonuçlar ile iki farklıgenin birlikte mikroenjeksiyonuyavruların transgenik olma yüzdelerinin yüksek olduğuortaya konmuştur. Sunulan çalışmada geliştirilenprotokolün transgenik fare üretim çalışmalarındauygulanması durumunda; iki farklı geni ayrı ayrı veyabirlikte edilebilecek, böylece ekonomi ve zaman açısındantasarruf sağlanabilecektir

Keywords:

Transgene, Micro-injection, EGFP, ,

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