【作者】 伍晓雄；

【导师】 李奎；

【作者基本信息】 华中农业大学 ， 动物遗传育种与繁殖， 2008， 博士

【摘要】 克隆出猪β-连环素基因片段,检测了该基因的组织表达谱。分析了不同群体中碱基突变位点的多态性,初步分析了该基因与部分生产性状和生理生化指标的关联性。对健康的通城（T）（n=33）、长白（L）（n=29）、大白（Y）（n=31）纯种猪以及长大通（LYT）（n=33）和大长通（YLT）（n=33）三元杂种猪的生理生化指标进行了对比研究。测定了五个群体猪血清总蛋白（TP）、白蛋白（ALB）、甘油三酯（TG）、总胆固醇（CHO）、尿素氮（BUN）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LCL-C）、肌酐（CRE）、K⁺、Cl^-、Ca²⁺、三碘甲状腺原氨酸（T3）、四碘甲状腺原氨酸（T4）、促甲状腺素（TSH）、胰岛素（INS）、瘦素（leptin）、神经肽Y（NPY）和胰岛素样生长因子-1（IGF-1）的含量,检测了通城猪与长白猪血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、乳酸脱氢酶（LDH）、肌酸激酶（CK）的活性和血糖含量。并将上述生理生化指标与五个群体猪的生长、胴体与肉质性状进行了关联分析。结果如下:1.猪CTNNB1基因的克隆、组织表达、多态性检测与性状关联分析（1）依据人cDNA信息,应用生物信息学及比较基因组学策略,克隆出猪β-连环素基因片段,并获得猪β-连环素基因cDNA编码区全长序列。（2）利用RT-PCR法,检测了成年长白母猪心、肝、脾、肺、肾、背肌、心肌、脂肪、胃、大肠、小肠、卵巢、子宫、胸腺和淋巴十五种组织中CTNNB1基因的表达。结果表明,CTNNB1基因的不同转录本在不同组织中均有表达,并存在表达差异。（3）在CTNNB1的第3内含子的162位（G-A转换）和第9内含子的64位（G-C转换）发现了两个潜在的突变位点。分析了不同群体中碱基突变位点的多态性,计算了该位点在7个猪品种中的基因型和基因频率,比较了各基因型在不同猪群的分布差异。研究发现,该位点在国内外猪群体中具有丰富多态性。（4）研究了CTNNB1基因第3内含子的162位点和第9内含子的64位点的多态性,初步分析了该基因多态位点与部分生产性状和生理生化指标的关联性。结果表明,不同基因型个体的PLBH,MS,TP,TSH和leptin存在显著差异（P＜0.05）。2.血清激素含量测定结果在五个群体中,大长通血清总T3显著高于长白猪（P＜0.05）。通城猪血清总T4极显著高于其它群体（P＜0.01）。大白猪血清TSH极显著高于通城猪、大长通和长大通。五个群体中血清胰岛素含量差异不显著（P＞0.05）。大白猪血清IGF-1显著高于其它群体。五个群体中,通城猪血清瘦素最高,长大通最低,两者差异显著（P＜0.05）。长大通血清NPY显著高于大白猪和通城猪。3.血浆蛋白、尿素氮和肌酐含量测定结果在五个群体中,通城猪血浆TP最高,与其它猪群体差异极显著（P＜0.01）。大白猪血浆ALB最高,与长白猪和长大通差异显著（P＜0.05）。通城猪血浆球蛋白浓度与大白猪、长大通、大长通和长白猪差异显著。4.血清脂类物质和血浆脂蛋白含量测定结果在五个群体中,通城猪血清CHO显著低于其它群体（P＜0.05）。通城猪与大白猪血清TG显著低于长白猪、大长通和长大通（P＜0.05）。长白猪血清HDL-C显著低于通城猪和大长通（P＜0.05）。长大通血清LDL-C显著高于长白猪（P＜0.05）。大白猪血清Cr极显著高于其它群体（P＜0.01）,通城猪极显著低于其它群体（P＜0.01）。5.血液生理生化指标与生长性状的相关分析结果在5个群体中,血清CK与日增重呈低度正相关（P＜0.01）。血清ALP与达上市日龄呈低度正相关（P＜0.01）。血清肌酐与初生重和日增重呈显著正相关（P＜0.01）,与达上市日龄呈显著负相关（P＜0.01）。血清IGF-1含量与日增重呈微弱正相关（P＜0.01）,与达上市体重日龄呈微弱负相关（P＜0.01）。血清BUN、Cl^-、Ca²⁺、T4和leptin与达上市日龄呈低度正相关（P＜0.01）,与日增重呈低度正相关（P＜0.01）。血清ALT与达上市日龄呈低度负相关（P＜0.05）,与日增重呈低度正相关（P＜0.01）。6.血液生理生化指标与胴体性状的相关分析结果血清T3、TSH、INS、NPY、TP、ALB、A/B、TG、HDL-C、LDL-C和K⁺与所有胴体性状呈微弱相关（|r|＜0.3000）。血清Leptin含量与内脂率呈低度正相关（P＜0.01）,与胴体斜长呈低度负相关（P＜0.01）,与板油率、平均膘厚呈微弱正相关。血清IGF-1与屠宰率、内脂率和6-7肋骨膘厚呈低度负相关（P＜0.01）,与板油率、眼肌面积等呈低度正相关（P＜0.01）。血浆球蛋白含量与内脂率呈低度正相关（P＜0.01）。血清BUN含量与屠宰率、胴体斜长和眼肌面积等呈低度正相关（P＜0.01）,与内脂率呈低度负相关（P＜0.01）。血清肌酐（Cr）含量与屠宰率、胴体长度和眼肌面积（LMA）呈显著正相关（P＜0.01）,与内脂率、板油率和膘厚呈负相关。血清钙离子与胴体长度和肋骨数呈低度负相关（P＜0.01）。7.血液生理生化指标与肉质性状的相关分析结果血清T4与MPV、MS、DL和IF呈负相关（P＜0.01）,与CS和WLP呈显著正相关（P＜0.01）。血清T3与MSF呈显著负相关（P＜0.01）,与IF呈显著正相关（P＜0.01）。血清TSH与MPV、MS、DL和IF呈显著正相关（P＜0.01）,与CS和WLP呈显著负相关（P＜0.01）。血清瘦素与MSF呈显著负相关（P＜0.01）。血清NPY含量与CS、WLP呈显著正相关（P＜0.01）,与MPV、MS、DL和IF呈显著负相关（P＜0.01）。血浆TP含量与CS呈高度正相关（P＜0.01）,与MS、DL、IF和MPV呈显著负相关（P＜0.01）。血浆ALB含量与MS、DL、MPV和IF呈显著负相关（P＜0.01）,与CS和WLP呈显著正相关（P＜0.01）。血液BUN含量与MS、DL、MPV和IF呈正相关（P＜0.01）,与CS和WLP呈显著负相关（P＜0.01）。血清LDL-C含量与DL、MS和IF呈显著正相关（P＜0.01）,与CS和WLP呈显著负相关（P＜0.01）。血清肌酐含量与MS、MPV、DL和IF呈负相关（P＜0.01）,与WLP和CS呈显著正相关（P＜0.01）。更多还原

【Abstract】 The coding region sequence of the porcineβ-catenin gene was cloned,and the tissue distribution of this gene was analyzed.Polymorphisms of mutational site in porcineβ-catenin gene were analyzed,and the association of the genotypes with some production traits and physiological and biochemical indexes was analyzed initially in 5 pig populations.The physiological and biochemical indexes in 5 healthy pig populations were studied in this research,including 3 purebreds of Tongcheng（n=33）,Large White（n=29）,Landraee（n=31）,and 2 hybrids of LYT （Landrace×（Large White×Tongcheng））（n=33） and YET（Large White×（Landrace×Tongcheng））.It were determined serum concentrations of total protein（TP）,albumin（ALB）,triglyceride（TG）,the total cholesterol（CHO）,blood urea nitrogen（BUN）,high density lipoprotein cholesterol（HDL-C）, low density lipoprotein cholesterol（LCL-C）,creatinine（CRE）,K⁺,Cl^-,Ca²⁺,triiodothyronine（T3）, tetraiodothyronine（T4）,thyrotropin（TSH）,insulin（INS）,leptin,neuropeptide Y（NPY） and insulin-like growth factor-1（IGF-1）.It was also determined the activities of serum alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,lacticdehydrogenase（LDH）, creatinkinase（CK） and the content of blood sugar in Tongcheng and Large White.Correlation analysis was conducted between physiological and biochemical indexes and the traits of growth,carcass and meat quality in 5 pig populations.The results as following:1.The results of cloning,tissue expression,polymorphisms detection and association analysis of the CTNNB1 gene（1） The full-length sequence of coding region of the porcineβ-catenin gene was cloned using the porcine specific primers which were designed from the porcine EST contig obtained by the method of comparative genomics according to the corresponding human cDNA sequence.（2） The tissue distribution was analyzed by RT-PCR in 15 different tissues of an adult female Landrace,including heart,liver,spleen,lung,kidney,dorsal muscles,the cardiac muscle,adipose tissue,stomach,large intestine,small intestine,oarium,uterus,thymus gland and lymph.The result showed that the CTNNB1 gene was expressed in all the tissues.（3） Two potential mutational sites were found in porcine CTNNB1 gene,one was at 162 site （G-A transformed） in intron 3,the other was at 64 site（G-C transformed） in intron 9.The polymorphisms for the two mutations was analyzed in 7 different pig breeds,and the SNP existed polymorohic in the breeds.（4） The RFLP of CTNNB1 gene at 162 site（G-A） in intron 3 and at 64 site（G-C） in intron 9 was analyzed in the 5 pig populations,including T,L,Y,LYT and YLT.The association of the genotypes with some production traits was analyzed.The preliminary results indicated that the mutation is associated with PLBH,MS,TP,TSH and leptin in different Allele genotype.2.The assaying results of serum hormonesThe concentrations of serum total T3（TT3） in LYT and YLT were higher than of in Landrace and Large White.The serum total T4（TT4） content of Tongcheng was remarkably higher than of the other 4 pig populations（P＜0.01）.The serum TSH concentration of Large white was remarkably higher than of Tongcheng,YLT and LYT.The serum IGF-1 concentration of Large white was significantly higher than Landrace,YLT,LYT,and Tongcheng.The serum Leptin content of Tongcheng was higher than of the other pig populations.The serum NPY content of LYT was significantly higher than of Large white and Tongcheng.3.The assaying results of plasma protein,urea nitrogen and creatinineThere was significantly difference in serum TP concentration between Tongcheng and the other populations.The serum ALB content of Large white was exceedingly higher than Landrace and LYT （P＜0.01）.The serum BUN content of Tongcheng was striking lower than of Landrace and LYT（P＜0.05）.4.The assaying results of serum lipides and plasma lipoproteinThe serum CHO level of Tongcheng was remarkably lower than of the other populations（P＜0.05）.The serum TG content of Landrace was higher than of Tongcheng and Large white.The serum TG level of Tongcheng and Large white were striking lower than of Landrace,YLT and LYT（P＜0.05）.The serum HDL-C concentrations of Landrace was significantly lower than of Tongcheng and YLT（P＜0.05）.The serum LDL-C level of LYT was notable higher than of Landrace（P＜0.05）.The serum Cr content of Large white was extremely higher than of the other populations（P＜0.01）,and the Cr level of Tongcheng was exceedingly lower than of the other populations（P＜0.01）.5.The correlation analysis results between physiological and biochemical indexes and growth traitsThere was low positive correlation between serum CK and average daily gain（P＜0.01）.There were significant positive correlation between serum creatinine and growth traits.There was weak positive correlation between serum IGF-1 and growth traits（P＜0.01）.There were low negative correlation between serum Cl^-,BUN,Ca²⁺,T4,leptin,ALT（P＜0.05） and growth traits（P＜0.01）.6.The correlation analysis results between physiological and biochemical indexes and carcass traitsThere were weak correlation between serum T3,TSH,INS,NPY,TP,ALB,A/B,TG,HDL-C, LDL-C,K⁺ and the all carcass traits.There were low positive correlation between serum leptin and PLCF（P＜0.01）,and low negative correlation with MICL（P＜0.01）.There were low negative correlation between serum IGF-1 and DP,PLCF,6-7 BF（P＜0.01）,and low positive correlation with PLF,10 BF,LMA,LLI,PH（P＜0.01）.It was low positive correlation between serum globulin and PLCF（P＜0.01）,and low negative correlation with NB（P＜0.01）.There were low positive correlation between serum BUN and DP,MICL,NBR,LMA,ALMA,PLBH（P＜0.01）,and low positive correlation with PLCF（P＜0.01）.There were significant positive correlation between serum creatinine and DP,MACL,MICL,NB,LMA,ALMA,SIL,PH,PLBH（P＜0.01）,and significant negative correlation with PLCF,PLF,6-7 BF,10 BF（P＜0.01）.There were low positive correlation between serum creatinine and ABF,SIL（P＜0.01）.There were low positive correlation between serum Ca²⁺ and MACL,MICL,NB（P＜0.01）.7.The correlation analysis results between physiological and biochemical indexes and meat quality traitsThere were highly negative correlation between serum T4 and marbling score（MS）,drip loss （DL） and intramuscular fat（IF）（P＜0.01）,and significant negative correlation with muscle pH value（MPV）（P＜0.01）,and significant positive correlation with color score（CS） and water lossing percentage（WLP）（P＜0.01）.It was significant negative correlation between T3 and MSF, and positive correlation with IF（P＜0.01）.There were highly positive correlation between serum TSH and MPV,MS,DL（P＜0.01）,and significant negative correlation with CS and WLP,and striking positive correlation with IF（P＜0.01）.It was significant negative correlation between Leptin and MSF（P＜0.01）.There were significant positive correlation between serum NPY and CS,WLP, and striking negative correlation with MPV,MS,DL and IF（P＜0.01）.There were highly negative correlation between serum TP and MS,DL,IF,and highly positive correlation with CS（P＜0.01）.There were negative correlation between serum ALB and MS,DL,MPV,IF,and significant positive correlation with CS and WLP（P＜0.01）.There were highly positive correlation between BUN and MS and DL（P＜0.01）,and significant negative correlation with CS,WLP,MPV and IF （P＜0.01）.There were highly negative correlation between serum creatinine and MS,DL and IF （P＜0.01）,and highly positive correlation with WLP(P＜0.01.更多还原