猪H-FABP基因第三内含子的遗传变异

以马身猪、大白猪、长白猪和杜洛克猪四个品种共190头猪为实验动物,成功扩增出猪H-FABP基因intron 3的全序列,全长1 350 bp,已向GenBank提交,检索号为DQ 002993。根据序列测定结果设计引物P2,扩增其中255bp的片段,应用PCR-SSCP分析发现P2-SSCP多态位点,该位点上具有两个等位基因A和B。在4个猪种中,AA基因型频率最高,A等位基因频率明显高于B等位基因频率。序列测定的结果表明,P2-SSCP的变异是由碱基A→G的替换造成的。     

猪H-FABP基因多态片段的序列分析

利用PCR-RFLP方法在猪H-FABP基因内确定了3个变异酶切位点,分别为5′-上游区HinfI-RFLP、内含子2的HaeIII-RFLP和HinfI*-RFLP（HinfI和HinfI*代表不同区域的HinfI酶切反应）。对每个多态片段进行克隆测序分析,结果表明,5′-上游区HinfI-RFLP是由于1324位的碱基T→C的突变引起;在内含子2中,HaeIII-RFLP变异酶切位点在1811位,发生了C到G的突变;HinfI*-RFLP是由于1970位发生T→C的碱基替换。 Abstract:Three variant restriction sites of porcine H-FABP gene,including HinfI-RFLP in 5′-upsream,HaeIII-RFLP and HinfI*-RFLP in intron 2,were confirmed by PCR-RFLP method.The polymorphic fragments were cloned and sequenced.The results revealed a single nucleotide substitution of T→C at position 1324 for HinfI-RFLP,C→G at position 1811 for HaeIII-RFLP and T→C at position 1970 for HinfI*-RFLP,respectively.     

西藏小型猪H-FABP基因的PCR-RFLP研究

目的研究西藏小型猪心脏脂肪酸结合蛋白（H-FABP）基因5’-上游区和第二内含子内的遗传变异。方法应用PCR-RFLP技术测定30头西藏小型猪H-FABP的基因型。结果（1）在5’-上游区的Hinf I-RFLP位点上,西藏小型猪表现出多态性,等位基因h的频率为0．80;（2）在在第二内含子内的Hae Ⅲ-RFLP位点上,西藏小型猪均为DD纯合子;（3）在第二内含子内的Hinf I＊-RFLP位点上,除一头猪表现为bb基因型外,其余猪都表现为BB基因型,等位基因B的频率为0．97;（4）除Hae Ⅲ-RFLP位点外,在其余位点上西藏小型猪均处于Hardy-weinberg不平衡状态。（5）在Hinf I-RFLP中,西藏小型猪表现为中度多态性（0．25〈PIC〈0．5）,而在其他位点的RFLP中表现为低度多态（PIC〈0．25）。结论可以利用西藏小型猪H-FABP基因5’-上游区的多态遗传标记来分析其与肌内脂肪的关系。     

山西马身猪及其杂交后代H-FABP基因的PCR-RFLP检测

运用PCR-RFLP法检测马身猪及其杂交后代H-FABP基因多态性。结果表明,马身猪在Hinf Ⅰ-RFLP位点上有HH和Hh两种基因型,在HaeⅢ-RFLP位点上只有DD基因型,在HinfⅠ*-RFLP位点上有BB和Bb两种基因型,等位基因以h、D、B占优势,其频率分别为0．958、1．000和0．968。其杂交后代在HaeⅢ-RFLP位点上只检测到两种基因型DD和Dd。     

鸡H-FABP基因多态性及其与屠宰性能的关联分析

以90日龄草科鸡为实验材料, 对H-FABP基因进行PCR扩增, 采用PCR-SSCP技术结合测序分析了H-FABP基因在草科鸡中的多态性。结果表明: 草科鸡中存在332 G→A、534 G→A、835 C→T、1131→A、1294C→A、2329C→T、2372C→T、2636C→T等8处突变。经过基因型与屠宰性能的关联分析得知: 在7对产生多态的引物中, 只有第1对引物的各基因型与活重、屠体重、胸肌重、腿肌重和半净膛重差异显著(P<0.05); 全净膛重在不同基因型间差异极显著(P<0.01)。 由此推测, H-FABP可能对于鸡屠宰性能具有很大的影响或与控制屠宰性能的主基因连锁。     

猪MyoG基因3′端PCR-SSCP遗传多态性及其遗传效应(英文)

采用PCR-SSCP方法对长白猪、大白猪、杜洛克猪、山西黑猪和马身猪共636头猪的肌细胞生成素(Myogenin,简称MyoG)基因3′端的遗传多态性进行检测,分析MyoG基因对猪的初生体质量、断奶体质量、6月龄体质量和背膘厚的影响。根据已发表的猪MyoG基因3′端侧翼序列设计3对引物,发现F1/R1引物对扩增的片段有多态性。统计结果发现:长白、大白、杜洛克猪种B基因为优势基因,其基因频率分别为0.8807、0.7256和0.8581;山西黑猪种A基因为优势基因,其基因频率为0.9359;马身猪种只检测到A基因。χ2独立性检验表明,基因型分布在外来猪种(长白猪、大白猪、杜洛克猪)与地方猪种(山西黑猪、马身猪)间存在极显著差异(P<0.01)。固定效应模型分析结果表明,初生体质量基因型间差异显著(P<0.05),而断奶体质量、6月龄体质量和背膘厚基因型间差异不显著(P>0.05)。最小二乘分析结果表明,BB基因型与其它2种基因型比较有较大的初生质量,同AA和AB型比较差异极显著(P<0.01)。因此,推测MyoG基因对个体的初生体质量存在一定的影响,选择带有B等位基因的个体有望提高个体的初生体质量。     

猪H-FABP基因多态性与肌内脂肪含量的关联分析

以山西白猪为实验材料,采用索氏脂肪抽提法对其背最长肌的肌内脂肪含量进行了测定。根据心脏脂肪酸结合蛋白(heart fatty acid-binding protein,H-FABP)基因的PCR-RFLP多态性,应用SPSS软件,用最小二乘法拟合一般线性模型(GLM)分析了H-FABP基因的不同位点的基因型对肌内脂肪含量的影响。结果表明,基因型hh、dd、BB对肌内脂肪含量影响的效应值最大。证明H-FABP基因可以作为肌内脂肪含量的侯选基因。     

大绿蛙Sox基因和Dmrt基因的PCR-SSCP分析

Sox基因家族与Dmrt基因家族在胚胎发育及性别分化中起着重要作用。本文采用PCR方法,扩增了大绿蛙Sox基因和Dmn基因的保守区,分别获得长约220bp和150bp的片段;SSCP分析结果显示大绿蛙Sox基因、Dmrt基因雌雄个体片段无差异,与人的有较大差异。本研究为探讨大绿蛙的性别决定机制及Sox基因与Dmrt基因的进化提供了分子资料。     

两种蛙Sox基因的PCR-SSCP分析

采用PCR技术,以参考人SRI,基因HMG-box的保守区序列而设计一对特异引物,扩增了黑斑蛙和金线蛙的Sox基因。结果两种蛙的雌雄个体均扩增出217bp的基因片段,与人对照相同。对扩增产物进行SSCP分析显示,两种蛙雌雄个体间的单链迁移率相同,两种蛙之间差异较小,而与人有较大差异。测序表明,两种蛙的Sox序列之间以及与各类物种的Sox基因都有非常高的相似性,充分显示出Sox基因在系统进化上的高度保守性。     

绵羊H-FABP基因单核苷酸多态性的研究

以小尾寒羊(48只)、宁夏滩羊(121只)、滩寒杂交羊F1(23只)、无角陶塞特(48只)、萨福克(24只)5个绵羊群体为实验材料, 利用PCR-SSCP和DNA测序技术对心脏型脂肪酸结合蛋白(H-FABP)基因(GenBank登录号: AY157617)外显子2和内含子2部分序列进行单核苷酸多态性(SNPs)检测及遗传多态性分析。结果表明: (1)引物2的PCR扩增产物中存在981(G/A)、1014(A/C)、1019(T/C) 和1058 (-/G ) 4个SNP位点, 表现为AA、BB、CC、AB、AC、BC、AD、CD和BD 9种基因型, 其中AA为优势基因型。经χ2检验后, 除滩羊和萨福克羊外, 其他群体的基因频率和基因型频率均处于Hardy-Weinberg平衡状态。群体遗传多态性分析表明: 宁夏滩羊、小尾寒羊和无角陶塞特羊3个群体中的多态信息含量(PIC)均处于0.25和0.50之间, 为中度多态, 萨福克羊和滩寒杂交羊F1为低度多态(PIC<0.25), 表明脂肪酸结合蛋白基因在不同绵羊品种中具有单核苷酸多态性, 可以进一步作为候选基因来分析其与肌内脂肪含量性状的关联性。(2)引物4的PCR扩增产物中检测到1个SNP多态位点为2407(T/C), 表现为HH、Hh和 hh 3种基因型, 基因型频率大小为HH＞Hh＞hh, 经χ2检验后, 在滩羊和无角陶塞特羊中均为达到Hardy-Weinberg平衡状态, 其多态信息含量均为低度多态(PIC<0.25), 而在小尾寒羊、滩寒杂交羊F1和萨福克羊均没有多态出现。     

Cloning and Tissue Expression of Chicken Heart Fatty Acid-Binding Protein and Intestine Fatty Acid-Binding Protein Genes

Fatty acid-binding proteins (FABPs) are members of a superfamily of lipid-binding proteins, occurring intracellularly in invertebrates and vertebrates. This study was designed to clone and characterize the genes of heart fatty acid-binding protein and intestine fatty acid-binding protein in the chicken. PCR primers were designed according to the chicken EST sequences to amplify cDNA of H-FABP and I-FABP genes from chicken heart and intestinal tissues. Analysis of sequence showed that the cDNA of the chicken H-FABP gene is 75 to 77% homologues to human, mouse, and pig H-FABP genes, and the chicken I-FABP gene is 71 to 72% homologues to human, mouse, and pig I-FABP genes. In addition, Northern blot analysis indicated that of the two genes, similar to the copartner of the mammal, H-FABP gene was expressed in a wide variety of tissues, and I-FABP gene was expressed only in intestinal tissues. The expression levels of the chicken H-FABP mRNA in heart and I-FABP mRNA in intestine had significant differences between the broilers from fat line and Bai'er layers at six weeks of age. The results of this study provided basic molecular information for studying the role of two FABPs in the regulation of fatty acid metabolism in avian species.     

Polymorphism of Heart Fatty Acid-Binding Protein Gene Associatied with Fatness Traits in the Chicken

Fatty acid-binding proteins (FABP) belong to a superfamily of lipid binding proteins that exhibit a high affinity for long chain fatty acids and appear to function in metabolism and intracellular transportation of lipids. The current study was designed to investigate the effects of heart (H)-FABP gene on chicken growth and body composition traits. The Northeast Agricultural University divergent broiler lines for abdominal fat and a broiler X silkie F₂ population were used in this study. Body weight and body composition traits were measured in the populations. Primers were designed according to the chicken H-FABP gene sequence. Polymorphisms between parental lines were detected by DNA sequencing. PCR-RFLP and PCR-fragment length polymorphism methods were developed to genotype the populations. The results showed that the H-FABP gene polymorphisms in the two populations were associated with abdominal fat percentage. It implied that H-FABP gene can be a candidate locus or linked to a major gene(s) that affects abdominal fat content in the chicken.     

中外十个猪种H-FABP基因遗传变异的研究

利用PCR-RFLP技术（限制性内切酶采用HinfI,HaeⅢ和Msp I）检测了杜洛克、长白、大约克、南昌白、二花脸、梅山、玉山黑猪、乐平花猪、金华两头乌及上高两头乌10个猪种共计561头猪在心肌脂肪酸结合蛋白（H-FABP）基因位点的遗传变异。结果表明：（1）在HinfI-RFLP位点上,南昌白猪均为HH纯合子,而其他猪种在此位点上均存在变异,外来品种以纯合子HH居多。（2）在HaeⅢ-RFLP位点上,外来猪种杜、长、大及培育品种南昌白猪均在该位点表现为多态,但中国6个地方猪种在该位点均无变异;（3）在Msp I-RFLP位点上,仅有杜洛克在该位点表现为多态。而所有检测的地方猪种均表现为AADD-型。     

Association Between PCR-SSCP of Bone Morphogenetic Protein 15 Gene and Prolificacy in Jining Grey Goats

The Jining Grey is a prolific local goat breed in P.R. China. Bone morphogenetic protein 15 (BMP15) gene that controls high fecundity of Inverdale, Hanna, Lacaune, Belclare, Cambridge, and Small Tailed Han ewes was studied as a candidate gene for the prolificacy of Jining Grey goats. According to the sequence of ovine BMP15 gene, six pairs of primers were designed to detect single nucleotide polymorphisms in exon 1 and exon 2 of the BMP15 gene in both high fecundity breed (Jining Grey goats) and low fecundity breeds (Boer, Liaoning Cashmere, and Inner Mongolia Cashmere goats) by single strand conformation polymorphism (SSCP). Two pairs of primers (F1/R1 and F2/R2) were used to amplify the exon 1. Four pairs of primers (F3/R3, F4/R4, F5/R5, and F6/R6) were used to amplify the exon 2. Only the products amplified by primer F5/R5 displayed polymorphism. Results indicated that two genotypes (AA and AB) were detected in prolific Jining Grey goats and only one genotype (AA) was detected in low fecundity goat breeds. In Jining Grey goats frequencies of genotypes AA and AB were 0.10 and 0.90, respectively. Sequencing revealed two point mutations (G963A and G1050C) of BMP15 gene in the AB genotype in comparison to the AA genotype. In Jining Grey goats the heterozygous AB does had 1.13 (p < 0.01) kids more than the homozygous AA does. These results preliminarily showed that the BMP15 gene is either a major gene that influences the prolificacy of Jining Grey goats or a molecular genetic marker in close linkage with such a gene.     

猪MyoG基因3'端PCR-SSCP遗传多态性及其遗传效应

采用PCR—SSCP方法对长白猪、大白猪、杜洛克猪、山西黑猪和马身猪共636头猪的肌细胞生成素（Myogenin,简称MyoG）基因3’端的遗传多态性进行检测,分析MyoG基因对猪的初生体质量、断奶体质量、6月龄体质量和背膘厚的影响。根据已发表的猪MyoG基因3’端侧翼序列设计3对引物,发现F1／R1引物对扩增的片段有多态性。统计结果发现：长白、大白、杜洛克猪种B基因为优势基因,其基因频率分别为0．8807、0．7256和0．8581：山西黑猪种4基因为优势基因,其基因频率为0．9359;马身猪种只检测到4基因。χ^2独立性检验表明,基因型分布在外来猪种（长白猪、大白猪、杜洛克猪）与地方猪种（山西黑猪、马身猪）间存在极显著差异（P〈0．01）。固定效应模型分析结果表明,初生体质量基因型闻差异显著（P〈0．05）,而断奶体质量、6月龄体质量和背膘厚基因型闻差异不显著（P〉0．05）。最小二乘分析结果表明,BB基因型与其它2种基因型比较有较大的初生质量,同似和AB型比较差异极显著（P〈0．01）。因此,推测MyoG基因对个体的初生体质量存在一足的影响,选择带有B等位基因的个体有望提高个体的初生体质量。