鸭脂肪酸结合蛋白基因的克隆和序列多态性分析

Cloning and Analysis of Polymorphism of Fatty Acid Binding Protein Gene in Ducks

根据鸡脂肪型脂肪酸结合蛋白(A-FABP)基因序列设计1对引物对鸭基因组进行PCR扩增,将扩增出的475 bp片段进行克隆和测序,序列分析表明该基因片段包含部分外显子2、3和完整的内含子2序列,与鸡、鹅、猪的A-FABP基因分别有84%、94%和77%的同源性。在此基础上又设计3对引物利用PCR-SSCP方法对获得的序列进行多态性研究。结果引物P3在樱桃谷鸭、山麻鸭等品种中发现4处单碱基突变,分别为:241处“A-T”突变;243处的“T-C”突变;258处的“T-C”突变;299处的“C-T”突变。这些点突变共产生了3种基因型,且基因型分布在各鸭品种间有显著差异。

A pairs of primers were designed according to the sequence of chicken adipocyte fatty acid of binding protein （A-FABP） gene, and then the A-FABP gene was amplified by PCR from duck genome. The product of 475bp length of PCR was cloned and sequenced, it includes partial exon 2, exon 3 and complete intron 2. Homologous comparison was done between duck and chicken, goose and porcine A-FABP gene and the homology are 84%, 94% and 77% respectively. Then 3 pairs of primers were designed on the base of the cloned gene, the SNPs was detected by the technique of single strand conformation polymorphism （SSCP） and then confirmed by sequencing. Four nucleotldes mutation were found, A-T at -241bp, T-C at -243bp, T-C at- 258bp and C-T at-299bp respectively. Three genotypes were detected based on these mutation, and genotypes frequencies among duck breeds were different significantly.