Identification of novel single nucleotide polymorphisms in the growth hormone ( GH ) gene in Anatolian water buffalo ( Bubalus bubalis ) populations in Turkey

ABSTRACT This study was conducted to investigate the growth hormone ( GH; somatotropin-like) gene polymorphisms in 150 water buffalo ( Bubalus bubalis ) from different regions of Turkey. 404 bp long partial intron 4, exon 5, 3’ UTR regions of the GH gene (also called GH/Alu I locus) and 347 bp long exon-intron 3 and partial exon 4 regions of the GH gene (also called GH/Msp I locus) were amplified, and their PCR products analyzed via DNA sequencing method. Seven genotypes due to twenty single nucleotide polymorphisms (SNP) and one deletion/insertion were identified in a 347 bp long region of the GH/Msp I locus. A missense mutation from glycine to glutamate amino acid and four silent mutations in the serine, threonine, and asparagine amino acids were determined in the exon 3 region of the GH gene. Four genotypes due to eight SNP were identified in a 404 bp long region of the GH/Alu I locus. A missense mutation from lysine to arginine amino acid and six silent mutations in Leucine, aspartate, histidine, lysine, arginine, and cysteine amino acids were revealed in the exon 5 region of the GH gene. The partial DNA sequence of the GH gene in water buffalos was reported, and these sequences were deposited at the NCBI Genbank database with the accession numbers MN266903-MN266909 and MN530973-MN530976. These SNP may have an effect on economic (such as body composition) and carcass traits, reproduction, and milk yield and content in water buffalo populations and may prove to be useful for water buffalo breeding.

摘要 本研究旨在探究土耳其不同地区150头水牛（Bubalus bubalis）的生长激素（growth hormone，GH，亦称类生长激素）基因多态性。本研究针对生长激素基因的两处片段开展扩增：其一为第4内含子部分区域、第5外显子及3’非翻译区（3’ untranslated region，3’UTR）的404 bp序列（亦称GH/Alu I位点）；其二为第3外显子-内含子区域与第4外显子部分区域的347 bp序列（亦称GH/Msp I位点）。随后通过DNA测序法对其聚合酶链式反应（polymerase chain reaction，PCR）产物进行分析。在GH/Msp I位点的347 bp区域内，共鉴定出由20个单核苷酸多态性（single nucleotide polymorphism，SNP）及1处插入/缺失突变所介导的7种基因型。在生长激素基因第3外显子区域中，检测到1处甘氨酸向谷氨酸的错义突变，以及丝氨酸、苏氨酸与天冬酰胺残基上的4处同义突变。在GH/Alu I位点的404 bp区域内，共鉴定出由8个SNP所介导的4种基因型。在生长激素基因第5外显子区域中，发现1处赖氨酸向精氨酸的错义突变，以及亮氨酸、天冬氨酸、组氨酸、赖氨酸、精氨酸与半胱氨酸残基上的6处同义突变。本研究已提交水牛生长激素基因的部分DNA序列，并将上述序列上传至NCBI GenBank数据库，登录号为MN266903-MN266909及MN530973-MN530976。上述SNP可能对水牛群体的经济性状（如体组成）、胴体性状、繁殖性能以及产奶量与乳成分产生影响，可为水牛育种工作提供有效参考。