Polymorphism analysis of h-fabp gene and correlation with imf content in wei pig | 圩猪H-FABP基因多态性分析及其与IMF含量的相关性

Chinese. 【目的】探究圩猪心脏脂肪酸结合蛋白(H-FABP)基因的多态性,及其与IMF含量的相关性【方法】采用PCR-RFLP(HinfⅠ、HaeⅢ及MspⅠ3种限制性内切酶)分子标记技术,分析了123头圩猪H-FABP基因5′-上游区和第二内含子区的遗传变异情况。【结果】(1)在5′-上游区：HinfⅠ位点上存在多态性,等位基因H的基因频率为0.6545,表现为中度多态(PIC=0.3500);第二内含子区：HinfⅠ*位点上存在多态性,等位基因B的基因频率为0.7195,表现为中度多态(PIC=0.3222);Hae Ⅲ位点上也存在多态性,等位基因D的基因频率为0.6301,表现为中度多态(PIC=0.3575);而MspⅠ位点上尚未检测到多态,基因型均为AA型。(2)被测猪群的基因频率和基因型频率都处于Hardy-Weinberg平衡状态(P＞0.05);(3)所检测的基因型对肌内脂肪(IMF)含量影响的趋势为：HH＞Hh＞hh,dd＞Dd＞DD,BB＞Bb＞bb;遗传效应值分别为：4.6918、4.2665、3.7712、4.6124、4.3167、3.8173、4.3042、4.2358、4.1970;(4)对H-FABP基因3个PCR-RFLP多态片段进行克隆测序分析发现,HinfⅠ-RFLP是由于1 324 bp处存在T→C的突变引起,HaeⅢ-RFLP是由于1 811 bp处存在C→G的突变引起,HinfⅠ*-RFLP是由于1 970 bp处存在C→T的突变引起。测序结果与GenBank公布的X98558和Y16180序列有97.9%及98.3%的同源性。【结论】圩猪H-FABP基因5′-上游区和第二内含子区的多态性对其IMF含量有一定影响,但是否可作为研究该基因与圩猪IMF含量相关的重要遗传标记还需做进一步研究。

English. 【Objective】 Polymorphism of heart fatty acid-binding protein (H-FABP) gene and its correlation with IMF content in Wei pig were studied. 【Method】 PCR-RFLP (HinfⅠ, HaeⅢ, MspⅠ3 kinds of restriction enzymes) and molecular marker techniques were used in the study and the genetic variation of H-FABP gene 5′-upstream region and second intron in 123 Wei pigs was analyzed. 【Result】 In 5′-upstream region, there was polymorphism in loci of the HinfⅠand the frequency of allele H was 0.6545, showing moderate polymorphism (PIC = 0.3500). In the second intron, polymorphism was detected in polymorphic loci HinfⅠ*, the frequency of allele B was 0.7195, showing moderate polymorphism (PIC = 0.3222). Loci of HaeⅢ also existed, allele D frequency was 0.6301, showing moderate polymorphism (PIC = 0.3575 ). While no polymorphism was detected in the loci of MspⅠ, which the genotype was AA genotype. The gene frequencies and genotype frequencies of pigs tested have reached the Hardy-Weinberg equilibrium (P＞0.05). The detection of the genotype of the IMF content had the trend of HH＞Hh＞hh, dd＞Dd＞DD, BB＞Bb＞bb, genetic effect values were as following: 4.6918, 4.2665, 3.7712, 4.6124, 4.3167, 3.8173, 4.3042, 4.2358, and 4.1970. Cloning and sequencing analysis on polymorphic fragments of three PCR-RFLP in H-FABP gene showed that HinfⅠ-RFLP is due to T→C mutation in 1 324 bp, HaeⅢ-RFLP is due to C→G mutation in 1 811 bp, HinfⅠ*- RFLP is due to C→T mutation in 1 970 bp. Sequencing results had 97.9% and 98.3% homology with the sequence X98558 and Y16180 published by GenBank. 【Conclusion】Polymorphisms in 5′-upstream region and the second intron of H-FABP gene had certain effect on IMF content in Wei pig. However, whether these polymorphisms are important genetic markers, which are related with H-FABP gene and IMF content in Wei pig, needs to be further studied.