圩猪H-FABP基因多态性分析及其与IMF含量的相关性

Polymorphism Analysis of H-FABP Gene and Correlation with IMF Content in Wei Pig

【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);HaeⅢ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。

【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（P0.05）.The detection of the genotype of the IMF content had the trend of HHHhhh,ddDdDD,BBBbbb,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.