Leptin (LEP) regulates the glucose homeostasis directly and centrally by the regulation of the insulin levels or indirectly by alternation of the levels of the other glucose metabolism regulator hormones. The present investigation studied the polymorphism in LEP gene which is related to fertility in 81 female Egyptian river buffalo. The PCR-RFLP pattern of the gene using the restriction enzyme Eco91I showed that all the animals had monomorphic pattern in the studied gene which consists of CC. A 511-bp fragment from LEP gene was amplified and sequenced. The homology between the amplified LEP gene fragment in buffalo and cattle, sheep, goat, human, and mouse on the nucleotides sequence level was 99, 97, 97, 87, and 79%, respectively, and on the translated amino acids sequence level was 100, 98, 98, 85, and 82%, respectively. Several SNPs were detected; among them, the T27C SNP disrupted an intronic splicing silencer. The A114G, A310G, G263A, and G379A SNPs disrupt exonic splicing enhancers, and the last two SNPs create new exonic splicing enhancers. The A114G, C163A, A211G, G288A, A310G, A322G, G330C, C348T, T360C, and G379A SNPs cause S71G, T87 N, N103S, E129K, E136G, Y140C, E143Q, R149W, S153P, and R159Q amino acids mutations. N103S, E129K, E136G, Y140C, E143Q, and S153P were classified as deleterious mutations. Y140, E143, N103, and R149 were the most conserved among the mutated amino acids. S71G only increased the stability of the leptin protein while the remaining mutations decreased it. Four SNPs were revealed among the tested animals. Twenty-one SNPs were found between the sequenced amplicon and the buffalo records in the Genbank. Some SNPs were predicted to have several effects on different biological processes like mRNA splicing, protein stability, and the gene functions.

中文翻译：

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埃及河水牛瘦素基因多态性检测并预测其在不同分子水平上的影响

瘦素（LEP）通过调节胰岛素水平直接集中调节葡萄糖稳态，或通过改变其他葡萄糖代谢调节激素的水平间接调节葡萄糖稳态。本研究对81头雌性埃及河水牛与生育力相关的LEP基因多态性进行了研究。使用限制性内切酶Eco91I对该基因进行PCR-RFLP图谱显示，所有动物所研究的基因均具有由CC组成的单态图谱。LEP 基因的 511 bp 片段被扩增并测序。水牛和牛、绵羊、山羊、人和小鼠中扩增的LEP基因片段在核苷酸序列水平上的同源性分别为99%、97%、97%、87%和79%，在翻译的氨基酸序列水平上同源性为分别为 100%、98%、98%、85% 和 82%。检测到多个 SNP；其中，T27C SNP 破坏了内含子剪接沉默子。A114G、A310G、G263A 和 G379A SNP 破坏外显子剪接增强子，最后两个 SNP 创建新的外显子剪接增强子。A114G、C163A、A211G、G288A、A310G、A322G、G330C、C348T、T360C 和 G379A SNP 导致 S71G、T87 N、N103S、E129K、E136G、Y140C、E143Q、R149W、S153P 和 R159 Q氨基酸突变。N103S、E129K、E136G、Y140C、E143Q 和 S153P 被归类为有害突变。Y140、E143、N103 和 R149 是突变氨基酸中最保守的。S71G仅增加了瘦素蛋白的稳定性，而其余突变则降低了它。在测试的动物中发现了四个 SNP。在测序的扩增子和 Genbank 中的水牛记录之间发现了 21 个 SNP。一些 SNP 预计会对不同的生物过程产生多种影响，如 mRNA 剪接、蛋白质稳定性和基因功能。