Pırlak Koyun Irkında Bir Batında Doğan Yavru Sayısı ile Ilişkili PRL ve BHR Genlerindeki inDel Varyasyonlar

Öz

Çiftlik hayvanı populasyonlarının moleküler seviyede analiz edilmesine olanak sağlayan çok sayıda moleküler genotipleme yöntemleri bulunmakla birlikte özgün oligonükleotidlerle yönlendirilmiş geleneksel Polimeraz Zincir Reaksiyonu (PZR) tekli genlerin incelenmesinde kullanılan hızlı ve uygun maliyetli bir tekniktir. Günümüze kadar koyunlarda bir batında doğan yavru sayısı üzerine major etkileri olan çok sayıda gen bildirilmiştir. DNA seviyesinde ekspresyon profilini şekillendiren bu genlerdeki varyasyonlar koyun ırklarında bir batında doğan yavru sayısında farklılıkların görülmesine neden olabilmektedir. Bu çalışma Pırlak koyun ırkında Prolaktin (PRL) intron 2 ve Büyüme Hormonu Reseptörü (BHR) intron 3 ve intron 4 gen bölgelerindeki insersiyon/delesyon (inDel) varyasyonlarının geleneksel PZR yöntemiyle incelendiği ilk araştırmadır. Antalya’da yetiştirilen, ırkı temsil eden ve birbiriyle akraba olmayan toplam 100 hayvan 23 baz çifti (bç) uzunluğundaki inDel’in varlığı/yokluğuna göre genotiplendirilmiş ve bütün varyasyonlar bakımından üç farklı genotip (II, ID ve DD) tespit edilmiştir. PRL-intron 2 lokusunda I ve D allel frekansı sırasıyla 0.421 ve 0.579 olarak bulunmuştur. BHR-intron 3 ve intron 4 lokuslarındaki I / D allel frekansları sırasıyla 0.599 / 0.401 ve 0.372 / 0.628 olarak bulunmuştur. En düşük II (0.181) ve DD (0.177) genotip frekansı sırasıyla BHR-intron 4 ve BHR-intron 3 lokusunda belirlenmiştir. Populasyon seviyesinde en düşük (DD için 0.177) ve en yüksek (ID için 0.448) genotip frekasnları BHR intron 3 lokusunda tespit edilmiştir. Sadece PRL-intron 2 lokusunda Hardy-Weinberg (HWD) dengesinden önemli sapma tespit edilmiştir. Bu çalışmanın sonuçları Pırlak ırkının PRL ve BHR gen bölgeleri bakımından yeterli seviyede genetik varyasyona sahip olduğunu ve bunun gelecekte bir batında doğan yavru sayısını arttırmak için yapılacak seleksiyon stratejilerinde kullanılabileceğini göstermiştir.

Anahtar Kelimeler

• Delesyon
• Insersiyon
• Bir batında doğan yavru sayısı
• Çoklu doğum
• PZR

InDel Variations of PRL and GHR Genes Associated with Litter Size in Pirlak sheep breed

Öz

Numerous molecular genotyping methods are available to analyse local livestock populations at molecular level in which traditional Polymerase Chain Reaction (PCR) guided by specific oligonucleotides is a fast and cost-effective method to investigate single genes. Until today, many genes which are of major effects on litter size have been reported in sheep. Genetic variations in these genes shaping the expression profile at DNA level may lead to differences in litter size among the sheep breeds. This is the first attempt to investigate insertion/deletion (inDel) variations in Prolactin (PRL) intron 2 and Growth Hormone Receptor (GHR) intron 3 and intron 4 genes in Pirlak sheep breed via traditional PCR technique. A total of 100 unrelated animals sampled from representative herds reared in Antalya were genotyped based on absence/presence of 23 base pairs (bp) length inDel in which three genotypes (II, ID, and DD) were detected in all loci. I and D allele frequency were 0.421 and 0.579, respectively in terms of PRL-intron 2 locus. I / D allele frequencies were found as 0.599 / 0.401 and 0.372 / 0.628 in GHR intron 3 and intron 4, respectively. The lowest II (0.181) and DD (0.177) genotype frequencies were detected in GHR-intron 4 and GHR-intron 3 loci, respectively. The lowest (0.177 for DD) and highest (0.448 for ID) genotype frequencies were detected in GHR intron 3 locus across the population. Significant deviation from Hardy-Weinberg Equilibrium (HWE) was detected only in PRL-intron 2 locus. The results of the present study confirm that Pirlak breed conserves sufficient genetic variation in PRL and GHR gene regions which could be utilized in selection strategies in order to increase litter size in the future.

Anahtar Kelimeler

• Deletion
• Insertion
• Litter size
• Multiple birth
• PCR

Kaynakça

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