藏绵羊ANGPTL4基因第3内含子、第6外显子多态性分析

Polymorphism of ANGPTL4 Gene in Tibet Sheep

为分析高寒牧区藏绵羊ANGPTL4基因多态性,应用PCR-SSCP与测序结合的方法,对391只藏绵羊ANGPTL4基因第3内含子、第6外显子进行多态性检测。经扩增,分别获得大小为280 bp和289 bp的2个片段。ANGPTL4基因第3内含子发现AA、BB和AB 3种基因型,各基因型频率分别为0.648 8、0.313 0和0.038 2,在C.400+90处发生G/T的突变,多态信息含量为0.265 6,呈中度多态。ANGPTL4基因第6外显子发现AA和AB2种基因型,基因型频率分别为0.961 8和0.038 2,在C.837处发生C/T的突变,为错义突变,并导致P.279处苏氨酸变为丝氨酸,多态信息含量为0.036 9,为低度多态。对藏绵羊ANGPTL4基因的多态性进行分析,发现的2处突变位点可作为藏绵羊品种改良以及种质资源保护的潜在分子标记。

In order to study the gene polymorphism ol Tibetan sheep ANGPTL4 gene,3 9 1copies ol Tibetan sheep blood samples were and collected their genomic DNA were extracted.The polymorphism ol intron-3 and exon- 6 ol the ANGPTL4 was tested by means ol PCR - SSCP combined with sequencing.By amplification two Iragments in size ol 2 8 0 bp and 2 8 9 bp were obtained.Primer amplilication products by the modilied polypro-pylene 3 belt types,namely AA,BB and AB,were lound Irom the intron- 3 ol ANGPTL4 gene, and their geno-type Irequencies were 0.648 8,0.313 0 and 0.038 2, respectively. The polymorphism information content was 0.265 6 , belonging to moderate polymorphism.Individual test showed that the primer amplilication lragment ol P 1 was 1 base mutation, G/T mutation was lound in c.400 G＋ 9 0 ;In primer amplilication ol P2 product by the modilied polypropylene amide gel electrophoresis, two types appeared, namely AA and AB, their genotype lre- quencies were 0.961 8 and 0.038 2, respectively.The polymorphism inlormation content was 0.036 9 belonging to low polymorphism.Individual test showed that the primer amplilication lragment ol P2 was 1 base mutation,C/T mutations occurred,which in C.83X/P .279 led in lead to the corresponding amino acid change.The ampli-fication loci found in this study can be used as potential molecular markers lor quality improvement and germ plasm protection of Tibetan sheep.