共查询到19条相似文献,搜索用时 93 毫秒
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《畜牧与兽医》2017,(3):9-13
为进一步探究脂蛋白脂酶(LPL)基因的功能,揭示其对从江香猪肉质性状的影响,本研究采用荧光定量RT-PCR技术结合酶活性检测方法分析了从江香猪不同组织LPL基因表达水平及活性规律。结果显示,LPL基因在从江香猪大脑、心脏、肝脏、脾脏、肺、肾脏、大肠、背肌、子宫和卵巢10种组织内均有表达;其中,背肌和心脏组织中表达量较高,子宫和肺组织中表达量较低,且背肌中LPL mRNA表达量显著高于其他各组织(P0.05)。从江香猪4种组织LPL活性分析结果表明,背肌组织中的LPL活性显著高于子宫和心脏组织(P0.05),说明背肌中脂肪沉积能力高于子宫和心脏组织,研究结果将为今后分析该基因对从江香猪肉质性能的影响奠定基础。 相似文献
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马身猪和大白猪不同组织DECR1基因的表达 总被引:1,自引:0,他引:1
旨在研究DECR1基因在猪不同组织和品种中mRNA和蛋白水平的表达规律,探讨该基因与脂肪代谢的关系。以山西马身猪与大白猪为试验材料,提取肝脏、心脏、肾脏、脾脏、肺脏、胃、小肠、皮下脂肪和背最长肌组织的总RNA和总蛋白,应用实时荧光定量PCR技术检测DECR1基因在2个品种各组织中mRNA的相对表达量,采用Western blot技术对各组织中DECR1蛋白进行半定量分析。实时荧光定量PCR结果显示:DECR1基因在各组织中均有表达,组织之间的表达量存在极显著差异(P<0.01),DECR1基因在肝脏、皮下脂肪与心脏中为高丰度表达;在不同品种的皮下脂肪组织中,大白猪DECR1的mRNA表达极显著高于马身猪(P<0.01)。Western blot检测结果显示:DECR1在各组织中均有表达,不同组织的表达量存在极显著差异(P<0.01),在皮下脂肪、肝脏与小肠中高表达;不同品种的皮下脂肪组织中,大白猪DECR1蛋白的表达显著高于马身猪(P<0.05)。猪DECR1基因在不同组织的表达差异可能与脂肪代谢和脂肪沉积有关。 相似文献
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试验旨在探明从江香猪β-干扰素(interferon-beta,IFN-β)基因编码区分子序列及原核表达产物特征。以从江香猪为研究对象,提取肝脏总RNA并反转录为cDNA,设计特异性引物扩增IFN-β基因编码区,将目的基因片段克隆至原核表达质粒pET-28a上,获得重组质粒pET28a-CJpoIFN-β,并利用生物学软件对江香猪IFN-β基因编码区进行序列分析;将鉴定正确的重组质粒pET28a-CJpoIFN-β转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导表达、SDS-PAGE与Western blotting分析原核表达蛋白。结果表明,从江香猪IFN-β基因编码区长为561bp,编码186个氨基酸;该蛋白为分泌性蛋白,前21个氨基酸为信号肽序列;二级结构主要以α-螺旋(77.42%)和无规则卷曲(17.74%)为主。从江香猪与其他猪源IFN-β基因核苷酸序列同源性为99.5%~100.0%,与禽的同源性最低(35.2%);从江香猪与巴马猪、梅山猪IFN-β氨基酸同源性均为100.0%,但与贵州白香猪IFN-β同源性为99.5%,存在E43Q、K73R和C161R3处氨基酸的差异。Western blotting结果显示,带His标签的重组表达蛋白能被His单抗识别,条带大小约为24ku。本试验结果为进一步研究IFN-β基因生物学活性及加快从江香猪这一品种资源的有效利用提供参考依据。 相似文献
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本研究以从江香猪为研究对象,二元杂交猪(从江香猪×野猪)、外三元杂交猪(杜×长×大)为对照,采用DNA池和直接测序技术对3个群体的SIM1基因7个外显子、部分内含子以及3'非翻译区序列进行多态性检测;利用生物信息学软件预测多态位点对SIM1基因mRNA二级结构和蛋白质一级、二级结构的影响。结果表明,在3个群体的SIM1基因中筛查到12个SNPs,C77T位于第5外显子,T29186C、A29195C位于第9内含子,C63T、C225T位于第10外显子,C107T、A426G、T583C、A586C、A605C、A615C位于第11外显子,G267T位于3'非翻译区。其中C77T、T29186C、A29195C、C63T、C225T、C107T、G267T为同义突变,A426G、T583C、A586C、A605C、A615C为错义突变;5个错义突变分别导致异亮氨酸(Ile)变为缬氨酸(Val)、亮氨酸(Leu)变为脯氨酸(Pro)、谷氨酸(Glu)变为丙氨酸(Ala)、谷氨酸(Glu)变为天冬氨酸(Asp)、天冬酰胺(Asn)变为组氨酸(His);根据在线软件预测,突变前后的SIM1基因mRNA二级结构和蛋白质一级、二级结构均会发生改变。 相似文献
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试验旨在探明从江香猪β-干扰素(interferon-beta,IFN-β)基因编码区分子序列及原核表达产物特征。以从江香猪为研究对象,提取肝脏总RNA并反转录为cDNA,设计特异性引物扩增IFN-β基因编码区,将目的基因片段克隆至原核表达质粒pET-28a上,获得重组质粒pET28a-CJpoIFN-β,并利用生物学软件对江香猪IFN-β基因编码区进行序列分析;将鉴定正确的重组质粒pET28a-CJpoIFN-β转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导表达、SDS-PAGE与Western blotting分析原核表达蛋白。结果表明,从江香猪IFN-β基因编码区长为561 bp,编码186个氨基酸;该蛋白为分泌性蛋白,前21个氨基酸为信号肽序列;二级结构主要以α-螺旋(77.42%)和无规则卷曲(17.74%)为主。从江香猪与其他猪源IFN-β基因核苷酸序列同源性为99.5%~100.0%,与禽的同源性最低(35.2%);从江香猪与巴马猪、梅山猪IFN-β氨基酸同源性均为100.0%,但与贵州白香猪IFN-β同源性为99.5%,存在E43Q、K73R和C161R 3处氨基酸的差异。Western blotting结果显示,带His标签的重组表达蛋白能被His单抗识别,条带大小约为24 ku。本试验结果为进一步研究IFN-β基因生物学活性及加快从江香猪这一品种资源的有效利用提供参考依据。 相似文献
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《中国兽医学报》2019,(7):1261-1268
干扰素(interfern,IFN)是在特定的诱生剂作用下,由细胞产生的一种具有高度生物活性的糖蛋白,具有广谱抗病毒活性。从江香猪源Ι型干扰素α2和Ⅱ型干扰素γ于生菜中融合表达及生物活性研究未见报道。本研究应用生菜植物表达系统,将从江香猪源干扰素植物表达载体PBI121-IFNα2、PBI121-IFNα2γ、PBI121-IFNγ、PBI121-Vec转入根癌农杆菌LBA4404中,用真空渗透法将携带干扰素质粒的根癌农杆菌感染生菜叶片;用β-葡萄糖苷酸酶(β-glucuronidase,GUS)染色和酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)检测从江香猪源干扰素在生菜叶片中的表达;用实时荧光定量PCR(quantitative real-time PCR,Q-PCR)技术分析从江香猪源干扰素蛋白活性。结果显示:GUS染色和ELISA检测证明从江香猪源IFNα2、IFNα2γ、IFNγ在生菜中实现了表达;Q-PCR检测显示:在Marc-145细胞上从江香猪源干扰素蛋白对PRRSV的复制有明显抑制作用,在PK15细胞上从江香猪源干扰素蛋白对干扰素γ诱导蛋白30(interferon-gamma-inducible protein 30,IFI30)、2′-5′寡聚腺苷酸合成酶(2′-5′oligoadenylate synthetase,OAS)、抗黏液病毒蛋白(myxovirus resistance,MX)基因的mRNA水平的上调有一定的促进作用。本研究成功实现从江香猪源IFNα2、IFNα2γ、IFNγ在生菜中进行表达且表达蛋白具有生物学活性,为从江香猪源Ι型和Ⅱ型干扰素新复合型药物植物蛋白的开发利用提供参考依据。 相似文献
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细胞凋亡可及时清除机体内多余和受损伤的细胞,以维持组织器官的稳定性。为探讨凋亡过程在公猪附睾发育过程中发挥的潜在作用,本实验以从江香猪为研究对象,利用实时荧光定量PCR(qRTPCR)、免疫组织化学(IHC)和WesternBlot技术分析了附睾15d(初情前期)、30d(初情期)、60d(初情后期)和180 d(性成熟期)4个时期凋亡相关因子Caspase-3、Bax和Bcl-2的差异表达模式。qRTPCR结果显示:Casp3和Bax基因mRNA在从江香猪60d表达量最高,Casp3在15d表达量最低,15d和180 d差异不显著;Bax在180 d表达量最低,且15 d、60 d和180 d差异不显著;Bcl-2在60 d表达量最低,在15 d表达量最高,4个日龄段差异显著。免疫组化结果显示:Caspase-3蛋白在30 d主要表达在附睾管腔的微绒毛,而在180 d主要表达在管腔中的精子上。Western Blot结果显示:Caspase-3蛋白表达丰度依次为60 d>30 d>15 d>180 d,且各日龄段差异显著。综上,本实验发现不同日龄从江香猪附睾凋亡相关... 相似文献
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MENG Lijie WANG Weiyong YANG Yi XU Yongjian FENG Xianzhou HUANG Yong GAO Yi GONG Ting 《畜牧兽医学报》1956,51(11):2720-2730
The aim of this experiment was to study the expression pattern of taste receptor family 1 subtypes 1 (T1R1) and 3 (T1R3) during epididymal development of Congjiang Xiang pig, and to explore the possible role of these taste receptors in mammalian male reproductive function and its potential medical value. In this study, the differential expressions of T1R1 and T1R3 in epididymis at 4 key developmental periods (neonatal (15 d), peri-puberty (30 d), puberty (60 d) and sexual maturity (180 d)) of Congjiang Xiang pigs were analyzed. RT-qPCR, immunohistochemistry (IHC) and Western blot were used to detect the changes and distribution of the two taste receptors in epididymis of Congjiang Xiang pigs at different ages. The results of RT-qPCR showed that the expression of TAS1R1 and TAS1R3 mRNA increased gradually from neonatal (15 d) to sexual maturity (180 d), and there was a significant difference between each period (P<0.01). The results of Western blot showed that the expression of T1R1/T1R3 protein was the highest on the 180 d and the lowest on the 15 d. The average protein abundance of T1R1/T1R3 was as follows: 180 d > 30 d > 60 d > 15 d. The results of IHC showed that T1R1 and T1R3 proteins were distributed in the epididymis of Congjiang Xiang pigs at 4 periods, in which T1R1 protein was mainly concentrated in epithelial cell membrane, especially in basal and narrow cells, while T1R3 protein was strongly positive in stereocilia, annular vacuoles and spermatozoa. In summary, the expression of T1R1/T1R3 in the epididymis of Congjiang Xiang pigs increased gradually from 15 d to the peak of sexual maturation, which was related to the differential expression of T1R1/T1R3 in epithelial basal cells, narrow cells and stereocilia of epididymis. These special expression patterns were time related to the physiological function of epididymis, so it is speculated that T1R1/T1R3 are involved in the regulation of sperm maturation and storage in epididymis. 相似文献
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T1R1和T1R3在从江香猪附睾发育中的表达模式 总被引:1,自引:0,他引:1
为研究味觉受体第一家族亚型1(T1R1)和3(T1R3)在从江香猪附睾发育过程中的表达模式,探讨味觉受体在哺乳动物雄性生殖机能中可能发挥的作用及潜在医学价值,本试验以从江香猪附睾组织为研究对象,分析附睾发育4个关键时期:初情前(15 d)、初情时(30 d)、初情后(60 d)和性成熟期(180 d)T1R1与T1R3的差异表达。采用实时荧光定量PCR、免疫组织化学(IHC)和Western blot检测两个味觉受体在不同日龄从江香猪附睾组织中转录、翻译水平的变化及其分布情况。RT-qPCR结果表明:TAS1R1与TAS1R3 mRNA在从江香猪附睾初情前(15 d)至性成熟期(180 d)表达量逐渐增加,且任意两个时期间差异极显著(P<0.01)。Western blot结果显示,T1R1/T1R3蛋白在180 d表达量最高,在15 d表达量最低,两者之间差异显著(P<0.05),平均表达丰度依次为180 d > 30 d > 60 d > 15 d。IHC结果显示,T1R1和T1R3蛋白在各日龄组从江香猪附睾组织均有分布,其中T1R1蛋白主要在上皮细胞膜上,尤其是基细胞和窄细胞;而T1R3蛋白主要在微绒毛、环状空泡和精子呈强阳性表达。综上,本研究发现不同日龄从江香猪附睾的T1R1/T1R3表达从15 d逐渐增加,至性成熟达到峰值,这一表达变化与附睾上皮基细胞和窄细胞及微绒毛的T1R1/T1R3的差异表达有关,这些特殊的表达模式与附睾生理功能存在时间关联,故推测T1R1和T1R3参与附睾内精子成熟和储存的调节过程。 相似文献
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ZHAO Jia-fu DUAN Zhi-qiang YANG Yuan-qing JI Xin-qin WANG Yuan-yuan SUN Cheng-juan 《中国畜牧兽医》2017,44(7):1954-1960
This experiment was aimed to clone apolipoprotein A1(ApoA1) gene of Congjiang Xiang pig, and study the subcelluar localiztion of ApoA1 gene in eukaryocyte. The recombination plasmid pEGFP-C1-ApoA1 was constructed with RT-PCR and other methods, and detected by colony PCR,double digestion and sequencing, after successful construction of the recombination plasmid pEGFP-C1-ApoA1,the subcellular localization of ApoA1 protein were analyzed by fluorescence co-localization technique in the 36 h-transfected HEK-293T cells. Compared with ApoA1 gene of Sus scrofa submission in GenBank, the results showed that six base mutations were found in ApoA1 gene of Congjiang Xiang pig, five of above mentioned mutations were sense mutations, causing alanine to glutamic acid, histidine to glutamine, valine to leucine and aspartic acid to glycine in 180,185,186 and 209 amino acid residues, respectively. Using PSOR Ⅱ Prediction and fluorescence co-localization, it was found that the expression of ApoA1 protein was observed mainly in the extracellular matrix (77.8%). In conclusion, ApoA1 gene of Congjiang Xiang pig was cloned successfully, and the expression of ApoA1 protein was mainly concentrated in the extracellular matrix. These results would provide a knowledge for further constructing the ApoA1 gene transgenic animal models, and contribute to understanding the relation between ApoA1 gene and the human obesity-induced diseases. 相似文献
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试验旨在克隆从江香猪载脂蛋白A1(apolipoprotein A1,ApoA1)基因,研究ApoA1基因在真核细胞中的亚细胞定位情况。通过提取从江香猪总RNA,采用RT-PCR、目的基因的连接、转化等方法构建携带有绿色荧光蛋白的pEGFP-C1-ApoA1重组质粒,并经菌落PCR、双酶切及测序鉴定正确后,转染HEK-293T细胞,36 h后观察荧光,分析ApoA1蛋白在真核细胞中的亚细胞定位情况。结果表明,从江香猪ApoA1基因与GenBank上公布的野猪序列相比,有6处发生了碱基突变,其中5处为有义突变,分别导致180位氨基酸由丙氨酸变为谷氨酸、185位氨基酸由组氨酸变为谷氨酰胺、186位氨基酸由缬氨酸变为亮氨酰胺、209位氨基酸由天冬氨酸变为甘氨酸;PSORT Ⅱ Prediction和荧光共定位试验结果均表明,ApoA1蛋白的表达主要集中在细胞外基质,约占总表达量的77.8%。本试验成功克隆了从江香猪ApoA1基因CDS区,且ApoA1蛋白的表达主要集中在细胞外基质中,为进一步构建ApoA1基因转基因动物模型、开展ApoA1基因与人类因肥胖引起的相关疾病关系的研究奠定基础。 相似文献
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试验旨在探讨从江香猪肌内前体脂肪细胞分化过程中相关基因的表达。采集3日龄从江香猪背最长肌,采用Ⅱ型胶原酶消化法分离肌内前体脂肪细胞,进行原代和传代培养,并对其进行形态学观察。诱导培养后,利用油红O染色法对其进行鉴定。采用实时荧光定量PCR方法检测细胞诱导分化0、24、48、72和144 h时脂肪相关基因丙酮酸脱氢酶激4(PDK4)、成纤维细胞生长因子10(FGF10)、脂联素(ADIPOQ)、脂肪酸合成酶(FAS)、脂蛋白脂酶(LPL)、CCAAT增强子结合蛋白α(C/EBPα)、脂肪细胞脂肪酸结合蛋白4(FABP4)、蛋白激酶B(AKT2)的表达,选择诱导0 h作为对照组。结果显示,分离的肌内前体脂肪细胞5 h开始贴壁,贴壁的细胞呈圆形,胞体透明,经传代后,细胞形态均一,经诱导培养后,油红染色呈红色。实时荧光定量PCR结果显示,PDK4、ADIPOQ、C/EBPα、FAS、FABP4和AKT2基因mRNA表达水平在诱导48 h时均呈现较高表达,极显著高于其余各阶段(P<0.01);FGF10基因mRNA表达水平在诱导24和48 h时均较高;LPL基因mRNA表达水平在诱导72 h时极显著高于对照组(P<0.01),之后明显下降;PDK4、ADIPOQ和FGF10基因mRNA表达水平在诱导144 h时均极显著低于对照组(P<0.01);C/EBPα基因mRNA表达水平在诱导144 h时显著高于对照组(P<0.05);FAS基因mRNA表达水平在诱导144 h时显著低于对照组(P<0.05);AKT2和LPL基因mRNA表达水平在诱导144 h时与对照组差异不显著(P>0.05)。本试验成功培养了从江香猪肌内前体脂肪细胞,并检测了不同诱导阶段脂肪相关基因的表达情况,为进一步研究从江香猪脂肪代谢和沉积提供参考依据。 相似文献
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中草药饲料添加剂对香猪生长性能和肉质的影响 总被引:2,自引:0,他引:2
研究了添加不同剂量的中草药饲料添加剂(0.3%和0.4%)对香猪生产性能和肌肉营养组分的影响。结果表明:0.3%和0.4%中草药组的日增重比对照组分别提高了42.98%(P<0.05)和30.06%(P<0.05);料重比分别降低了14.69%(P<0.05)和13.73%(P<0.05)。中草药添加组肉质的pH24值、失水率、粗脂肪、粗蛋白、眼肌面积均显著高于对照组(P<0.05);0.4%中草药组的主要鲜味氨基酸和必需氨基酸含量含量分别较对照组提高了28.37%和28.75%;人体必需脂肪酸中,0.3%和0.4%中草药组较对照组分别提高2.18%和4.36%;不饱和脂肪酸各组中油酸、亚油酸含量最高,0.3%和0.4%中草药组的亚麻酸、亚油酸和花生四烯酸含量分别比对照组提高2.41%、2.67%、3.68%和5.12%、3.51%、19.12%。 相似文献
18.
YANG Yang XU Hou-qiang CHEN Wei ZHOU Di XU Min ZHANG Qing-qing ZHAO Huan-ping SUN Cheng-juan WANG Yuan-yuan ZHANG Ming YANG Tao 《中国畜牧兽医》2017,44(12):3401-3409
The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition. 相似文献
19.
试验旨在克隆获得PDK4、FGF10基因,并研究大白猪与从江香猪不同组织中PDK4、FGF10基因mRNA的表达差异。采用RT-PCR分别克隆从江香猪PDK4、FGF10基因并进行生物信息学分析,利用实时荧光定量PCR技术检测PDK4、FGF10基因在大白猪和从江香猪不同组织中mRNA的相对表达量。结果显示,从江香猪PDK4基因的编码区全长1 224 bp,编码407个氨基酸;FGF10基因的编码区全长636 bp,编码211个氨基酸。经BLAST软件进行同源性比对,发现从江香猪PDK4基因与羊、马、人的核苷酸序列同源性分别为93%、92%和91%;FGF10基因与羊、牛、人、鼠的核苷酸序列同源性分别为94%、93%、93%和90%。由PDK4基因系统进化树可知,从江香猪与牛、绵羊亲缘关系较近;由FGF10基因系统进化树可知,从江香猪与绵羊、牛、人、猕猴亲缘关系较近,与小鼠和鸡亲缘关系较远。实时荧光定量PCR结果显示,在从江香猪不同组织中,PDK4基因在肾脏中的表达量最高,在胃和脂肪中表达量较高,FGF10基因在胃中表达量最高,在肾脏和脂肪中表达量较高,两个基因在背最长肌中的表达量均最低;在大白猪的不同组织中,PDK4、FGF10基因在脂肪中的表达量均最高,PDK4基因在背最长肌中的表达量最低,而FGF10基因在心脏中表达量最低。本试验成功克隆了从江香猪PDK4、FGF10基因,并检测了其在大白猪与从江香猪不同组织中的表达,为进一步研究PDK4、FGF10基因在脂质代谢及脂肪沉积等方面的调控作用提供科学依据。 相似文献