College of Animal Science and Technology, Northwest A&F University, No. 22 Xinong Road, Yangling, 712100, Shaanxi, People's Republic of China.
Mol Biol Rep. 2013 Feb;40(2):857-63. doi: 10.1007/s11033-012-2126-6. Epub 2012 Oct 16.
In this study, we cloned the full coding region of NGF gene from the caprine ovary. Result showed the caprine NGF cDNA (GenBank Accession No. JQ308184) contained a 726 bp open reading frame encoding a protein with 241 amino acid residues. Bioinformatic analysis indicated that caprine NGF amino acid sequence was 83-99 % identical to that of mouse, pig, dog, human and bovine. It was predicted that caprine NGF contained nine serine phosphorylation loci, four threonine phosphorylation loci and nine specific PKC phosphorylation loci. The NGF mRNA expression pattern showed that NGF gene was expressed highly in ovary. This work provided an important experimental basis for further research on the function of NGF in goat. A single nucleotide polymorphism (A705G) in the coding region of NGF gene was detected by PCR-RFLP and DNA sequencing in 630 goats of three breeds. The frequencies of G allele were 0.52-0.61, and frequencies of A allele were 0.48-0.39 for SN, GZ and BG breeds, respectively. The does with GG genotype had higher litter size than those with GA and AA genotypes (P < 0.05). Hence, the biochemical and physiological functions, together with the results obtained in our investigation, suggest that the NGF gene could serve as a genetic marker for litter size in goat breeding.
本研究从山羊卵巢中克隆了 NGF 基因的完整编码区。结果表明，山羊 NGF cDNA（GenBank 登录号：JQ308184）含有 726bp 的开放阅读框，编码 241 个氨基酸残基的蛋白质。生物信息学分析表明，山羊 NGF 氨基酸序列与小鼠、猪、狗、人、牛的氨基酸序列具有 83-99%的同源性。预测山羊 NGF 含有 9 个丝氨酸磷酸化位点、4 个苏氨酸磷酸化位点和 9 个 PKC 特异性磷酸化位点。NGF mRNA 表达模式表明 NGF 基因在卵巢中表达水平较高。本研究为进一步研究 NGF 在山羊中的功能提供了重要的实验依据。通过 PCR-RFLP 和 DNA 测序在三个品种的 630 只山羊中检测到 NGF 基因编码区的单核苷酸多态性（A705G）。SN、GZ 和 BG 品种的 G 等位基因频率分别为 0.52-0.61，A 等位基因频率分别为 0.48-0.39。具有 GG 基因型的母羊产仔数高于具有 GA 和 AA 基因型的母羊（P<0.05）。因此，生化和生理功能以及我们的研究结果表明，NGF 基因可以作为山羊产仔数的遗传标记。
