Characteristics of Intron 17 and Exon 18 of Leptin Receptor Gene in Nigerian Locally Adapted and Exotic Turkeys

Oguntade Do
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Abstract

Leptin Receptor gene (LEPR) gene has been implicated as the activator of the JAK-STAT signaling pathway and plays an important role in regulating body energy storage and metabolism. Identification of Single nucleotide polymorphisms (SNPs) were carried out in intron 17 and exon 18 of LEPR gene in 200 turkeys comprising 100 each of Nigerian locally adapted and exotic (Nicholas White) turkeys. Blood samples for DNA extraction were collected at 20 weeks. DNA extraction was carried out using whole blood collected with Quick- DNA Miniprep Plus extraction kits by strictly followed the manufacturer’s protocol. The sequence data obtained were analyzed using GeneScan and Sequencing software (Applied Biosystems). The single nucleotide polymorphisms (SNPs) present in LEPR gene of the two turkey genotypes were identified using DnaSP 6 software. The allele frequency of each SNP, Heterozygosity (He) and Polymorphic information content (PIC) of the SNPs were calculated. Out of six SNPs identified in intron 17 of the turkey LEPR gene, four were found in local turkeys while all the six were found in exotic turkey. The major allele frequency of the SNPs ranged from 0.86 to 0.96. The SNP 28C>G in exotic turkey had the highest HE while SNPs 7T>G, 67A>T and 85G>C in local turkey had the least. The PIC ranged from 0.077 in local turkey to 0.211 in exotic turkey. In addition, ten SNPs were identified in exon 18 of the turkey LEPR gene, out of which eight were detected in local turkeys while seven were detected in exotic turkeys. The major allele frequency of the SNPs identified in exon 18 ranged from 0.54 in 602A>C in local turkey to 1.00 in 60T>G, 267A>G, 274A>G alleles that are fixed in both local and exotic turkeys. Also, SNP 602A>C in local turkey had the highest HE while incidentally SNPs 130G>C, 166G>A and 789G>A also in local turkey had the least heterozygosity. PIC discovered in exon 18 of LEPR gene ranged from 0.077 to 0.377 in local turkey. Intron 17 and exon 18 of LEPR gene is polymorphic in both Nigerian locally adapted and exotic turkeys and therefore could be used for further analysis to form a basis for genetic variation in growth in the two genotypes
尼日利亚本地适应和外来火鸡瘦素受体基因17和18的内含子特征
瘦素受体基因(Leptin Receptor gene, LEPR)被认为是JAK-STAT信号通路的激活因子,在调节机体能量储存和代谢中起重要作用。对200只火鸡的LEPR基因内含子17和外显子18进行了单核苷酸多态性(snp)鉴定,其中尼日利亚本地适应火鸡和外来(尼古拉斯怀特)火鸡各100只。20周时采集血样进行DNA提取。DNA提取使用Quick- DNA Miniprep Plus提取试剂盒采集的全血,严格按照制造商的方案进行。使用GeneScan和测序软件(Applied Biosystems)分析获得的序列数据。利用dnasp6软件对两种火鸡基因型的LEPR基因进行单核苷酸多态性(snp)鉴定。计算每个SNP的等位基因频率、杂合度(He)和多态性信息含量(PIC)。在火鸡LEPR基因的内含子17中发现的6个snp中,有4个在本地火鸡中发现,而所有6个都在外来火鸡中发现。主要等位基因频率在0.86 ~ 0.96之间。外来火鸡的SNP 28C>G的HE最高,本地火鸡的SNP 7T>G、67A>T和85G>C的HE最低。PIC范围从本地火鸡的0.077到外来火鸡的0.211。此外,在火鸡LEPR基因的第18外显子中发现了10个snp,其中8个在本地火鸡中检测到,7个在外来火鸡中检测到。在18外显子上发现的主要等位基因频率范围从本地火鸡602A>C的0.54到本地火鸡和外来火鸡固定的60T>G、267A>G、274A>G等位基因的1.00。此外,地方火鸡的SNP 602A>C的HE最高,顺便说一下,地方火鸡的SNP 130G>C、166G>A和789G>A的杂合性也最低。在当地火鸡LEPR基因18外显子中发现的PIC范围为0.077 ~ 0.377。LEPR基因的17号内含子和18号外显子在尼日利亚本地和外来火鸡中都是多态的,因此可以用于进一步分析,形成两种基因型生长遗传变异的基础
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