Faruk BOZKAYA, ŞÜKRÜ GÜRLER, Mugdat YERTÜRK

Investigation on the polymorphism of some loci by using PCR-RFLP in Japanese Quails (Coturnix coturnix japonica) raised in different locations of Turkey

Bu çalışmanın amacı Japon bıldırcınlarında (Coturnix coturnix japonica) 1. (CJA1) ve 3. (CJA3) kromozom üzerinde bulunan bazı genlerdeki kesim bölgesi polimorfizminin (RFLP) araştırılmasıdır. Bu çalışma ile rekombinasyon oranları ile ilgili araştırma amacıyla kullanılacak bir popülasyonu oluşturacak bireylerin seçimine temel teşkil edecek olan popülasyon genetiği parametrelerinin ortaya konması amaçlanmıştır. Türkiyenin altı farklı ilindeki işletmelerden elde edilen döllü bıldırcın yumurtaları 37°Cde 120 saat süreyle inkube edildikten sonra 191 adet embriyodan doku örneği alınmıştır. Alınan doku örneklerinden özel kitler yardımıyla DNA izolasyonu yapılmıştır. SEMA3E, IFR1, HAL, LOC396025, UGP2, LOC396192, TLX ve BMP5 lokuslarının belirli bölgeleri özel olarak dizayn edilmiş olan primerler yardımıyla ve PCR işlemi ile çoğaltılmıştır. Elde edilen PCR ürünleri uygun bir restririksiyon enzimi ile kesilmiş ve agaroz jel elektroforezi yardımıyla ayrımlanmıştır. İncelenen popülasyonlarda farklı allellerin varlığı, genotip ve allel frekansları, heterozigotluk dereceleri ve genetik mesafeler hesaplanmıştır. Çalışmada kullanılan sekiz lokustan 1. ve 3. kromozomlar üzerinde bulunan SEMA3E ve TLX lokuslarında polimorfizm tespit edilmiş, beş lokus monomorfik olarak bulunmuş bir lokusta (HAL) ise PCR işlemi ile çoğaltma yapılamamıştır. Çalışılan popülasyonların büyük oranda Hardy-Weinberg dengesinde oldukları gözlenmiştir. Araştırmanın sonuçları SEMA3E ve TLX lokuslarının çalışılan popülasyonlarda rekombinasyon oranlarının araştırılması amacıyla kullanılabileceğini göstermiştir.

Türkiyedeki Japon bıldırcınlarında (Coturnix coturnix japonica) bazı lokuslardaki polimorfizmin PCR-RFLP yöntemi ile araştırılması

The aim of this study was to investigate the restriction fragment length polymorphism (RFLP) of some loci on chromosomes 1 (CJA1) and 3 (CJA3) in Japanese quails (Coturnix coturnix japonica) raised in Turkey. With this study population genetics parameters were estimated in order to select individuals for establishing a reference population, which would be used for studies on recombination frequency. Fertilized eggs obtained from flocks raised in six different provinces of Turkey were incubated for 120 h and 191 embryos were collected. From the tissue samples of the embryos, DNA was isolated by using DNA isolation kits. Particular regions of SEMA3E, IFR1, HAL, LOC396025, UGP2, LOC396192, TLX and BMP5 loci were amplified with specifically designed primers for each locus by using PCR technique. The PCR products were cut with an appropriate restriction enzyme for each locus and analysed by using agarose gel electrophoresis. Presence of different alleles, allele and genotype frequencies, heterozygosities and genetic distances were estimated. Out of the eight loci studied, polymorphism was found for the SEMA3E and TLX loci on 1. and 3. chromosomes, respectively, while five loci were found to be monomorphic and one locus (HAL) could not be amplified by PCR. The populations studied were found to be mostly in Hardy-Weinberg equilibrium. The results indicated that the SEMA3E and TLX loci can be used for studying recombination frequencies in the populations included into the study.

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