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Christophe Tastet Florence Val Michel Lesage Lionel Renault Laurent Marché Michel Bossis Didier Mugniéry 《European journal of plant pathology / European Foundation for Plant Pathology》2001,107(8):821-832
Two major proteins, Mcf-A67 and Mcf-B66, were identified by mini two-dimensional polyacrylamide gel electrophoresis in order to distinguish the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from eight other species. These quarantine proteinic markers have been microsequenced after enzymatic digestion. The internal amino acid sequences exhibit similarities to members of a family of low molecular weight intracellular lipid-binding proteins. Moreover, to explore a simple, rapid, and inexpensive way to identify the two quarantine nematodes, dot blot hybridizations were performed using an antiserum (A67) produced from the longest amino-acid sequence of the protein Mcf-A67. Although several proteins stained on the M. chitwoodi and M. fallax western blot membranes, the two nematodes were easily distinguished from other root-knot nematodes, on dot blot assays with soluble proteins extracted from a single female. Because of its specificity and sensitivity, the use of the A67 antiserum to improve the diagnosis of the two European quarantine root-knot nematodes is discussed. 相似文献
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B.E. TORSTENSEN D.A. NANTON P.A. OLSVIK H. SUNDVOLD & I. STUBHAUG 《Aquaculture Nutrition》2009,15(4):440-451
Relative gene expression pattern of fatty acid transport proteins (FATP and cd36), intracellular fatty acid-binding proteins (FABP3, FABP10 and FABP11), β-oxidation-related genes [carnitine palmitoyl transferase II (CPTII), peroxisome proliferator-activated receptor β (PPARβ), acyl-CoA oxidase (AOX), long-chain fatty acyl-CoA synthetase (FACS), acyl-CoA dehydrogenase (dehydrogenase)] and uncoupling protein 2 (UCP2) was assessed by RT-qPCR in Atlantic salmon muscle (red and white), liver, heart, myosepta and visceral fat. FABP11, a FABP isoform not previously described in Atlantic salmon, was highly expressed in visceral fat and myosepta and at the lower level in red muscle, white muscle, myosepta and heart. Furthermore, Atlantic salmon were fed either a diet containing fish oil (FO) or a complete replacement of FO with a vegetable oil blend (55% rapeseed oil, 30% palm oil and 15% linseed oil; VO) for the production cycle (27 months from start of feeding and until ∼4.5 kg mean weight). The expression of genes related to β-oxidation, fatty acid uptake and transport in the white muscle indicate ( n = 3) significant down-regulation in VO fed Atlantic salmon and correlated with previously reported white muscle triacylglycerol stores and β-oxidation. FABP11 in visceral fat and myosepta was also down-regulated in VO fed fish. 相似文献
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脂肪型脂肪酸结合蛋白（adipocyte fatty-acid binding protein,A-FABP）在家畜脂肪沉积中发挥重要作用,但在藏绵羊群体中的遗传特性和特异的作用机制尚不明确,研究不同生产方向绵羊品种的FABP4基因变异及分布有助于揭示藏绵羊独特的种质特性。应用PCR-SSCP（polymerase chain reaction-single strand conformation polymerase）和测序技术检测了中国和新西兰共10个绵羊群体（575只个体）FABP4基因编码区（外显子2和外显子3）的变异,并分析其连锁不平衡状态。结果表明,两多态区域共鉴定8处单核苷酸突变,两多态区域SNPs间呈弱连锁不平衡状态（D′=0.548,r2=0.119）,鉴定14种潜在的单体型。外显子2-内含子2区域A1为优势等位基因（38.4%）,A1B1为优势基因型;外显子3-内含子3区域B2为优势等位基因（48.3%）,A2B2为优势基因型。新西兰绵羊群体在两多态区域偏离Hardy-Weinberg平衡状态（P<0.01）,而藏绵羊处于Hardy-Weinberg平衡状态（P>0.05）。罗姆尼与派仑代群体基因型分布无显著差异（P>0.05）,美利奴与考力代群体基因型分布差异显著（P<0.05）。新西兰绵羊群体两多态区域均为高杂合度及高度多态,藏绵羊群体均为高纯合度及中度多态。聚类分析表明欧拉和甘伽羊、考力代和美利奴羊、罗姆尼和派仑代羊分别聚在一起。c.246+37A>G和c.348+298T>C位点多态性在藏绵羊和新西兰绵羊群体中分布差异较大,该两处SNPs可作为潜在的分子标记应用于藏绵羊肌内脂肪含量性状选育。 相似文献
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秦川牛FABP4基因重组腺病毒载体的构建与鉴定 总被引：1，自引：1，他引：0
【目的】旨在通过构建秦川牛FABP4基因的重组腺病毒载体,为在细胞水平研究FABP4基因的功能和作用机制做准备。【方法】根据GenBank收录的牛FABP4基因mRNA序列设计引物,PCR扩增并克隆测序。将目的基因连接到穿梭载体pAdTrack-CMV上并用PmeⅠ线性化后,转化含有腺病毒骨架载体pAdEasy-1的E. coli BJ5183感受态细胞进行同源重组,得到重组质粒pAd-FABP4,并用PacⅠ酶切鉴定。将经过PacⅠ线性化的pAd-FABP4转染HEK 293A细胞进行病毒包装和扩增,TCID50法测定病毒滴度。【结果】经测序鉴定本次克隆的FABP4 序列与GenBank收录的一致。 酶切鉴定、绿色荧光观察、PCR及测序检测均证明,重组腺病毒载体构建成功并获得重组腺病毒AD-FABP4,病毒滴度为1.58×109 PFU•mL-1。【结论】成功构建了秦川牛FABP4基因重组腺病毒载体并获得高滴度的重组腺病毒。 相似文献
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为深入了解棉铃虫Helicoverpa armigera脂肪酸结合蛋白HaFABP1在参与细胞内脂肪酸代谢过程中的功能,基于酵母单杂交结果从幼虫中肠cDNA中扩增得到HaFABP1的开发阅读框,构建重组菌株BL21-pET28a-HaFABP1,进行融合蛋白His-HaFABP1的诱导和纯化,并利用实时荧光定量PCR检测棉铃虫不同时期和组织中HaFABP1的表达规律,最后检测2-十三烷酮处理下棉铃虫6龄幼虫中肠组织内HaFABP1的变化情况。结果显示,克隆获得的棉铃虫HaFABP1为405 bp,编码134个氨基酸,相对分子量和等电点分别为15.08 kD和5.54;在37℃下用0.5 mmol/L异丙基硫代半乳糖苷(IPTG)诱导重组菌株4 h后,得到约18.4 kD的可溶性蛋白His-HaFABP1,纯化后的融合蛋白条带单一且大小符合预期;HaFABP1在棉铃虫的脂肪体、中肠、体壁和头部中都有表达,在头部的表达量最低,在中肠中的表达量最高;HaFABP1在棉铃虫幼虫的每个发育时期都有表达,表达量总体呈现先降低后升高的趋势,4龄时最低而1龄时最高;10 mg/g 2-十三烷酮处理饲料饲喂棉铃虫6龄幼虫6 h和15 h后,中肠内HaFABP1的表达量与对照相比显著提高,且随着时间的延长逐渐增加。表明HaFABP1与棉铃虫的生长发育有关,并且有可能参与了昆虫的解毒过程。 相似文献
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To investigate the role of FABP4 on regulating fatty acid metabolism and autophagy in macrophage infected with BCG, small interfering RNA for FABP4 were transfected into RAW264.7 cells alone or combined with BCG infection. The immunoblot and immunofluorescence were used to detect the expression of FABP4, the accumulation of lipid droplets, the expression of fatty acid β-oxidation and autophagy related factors. The results showed that the infection of BCG increased the expression of FABP4 and accompanied with the accumulation of lipid droplets in RAW264.7. Moreover, the knockdown of FABP4 decreased lipid droplets but promoted the expression of carnitine palmitoyltransferase 1A (CPT1A) (P <0.05) and the production of ATP (P <0.05). Meanwhile, the knockdown of FABP4 suppressed the expression of AMPK, p-ULK1, ATG5, ATG7, ATG12 and LC3B which are autophagy related factors (P<0.001). Our results indicated that the knockdown of FABP4 promoted fatty acid oxidation, decreased fatty acid level and suppressed autophagy via AMPK signal pathway in BCG infected RAW264.7 cells. 相似文献
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The study of CCAAT/enhancer binding proteins (C/EBPs) is important in the understanding of adipogenesis, but little is known about their regulation in fish. Here, we report three Atlantic salmon orthologs of c/ebp , and their expression in different tissues and in adipogenesis in vitro . During differentiation the expression of c/ebp α and fatp1 were upregulated in early differentiation stage with continuing high expression level in mature adipocytes, whereas c/ebp δ and fabp11 expression were elevated in mature adipocytes. Furthermore, the fatty acids eicosapentaenoic (EPA) and docosahexaenoic (DHA), suppressed the expression of the c/ebps, ppar β , and fatty acid transport protein ( fatp1 ) during terminal adipocyte differentiation. The study indicates that C/EBPs are induced upon the differentiation of primary-cultured adipocytes from Atlantic salmon and that marine n-3 highly unsaturated fatty acids (HUFAs) affect the c/ebp s expressions in mature adipocytes. Therefore, the established cell model described here appears to be valuable for studying modulation of fat content in farmed Atlantic salmon. 相似文献
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ABSTRACT 1. Fatty acid-binding proteins (FABP) are members of lipid-binding proteins, which participate in the metabolism and intracellular transportation of lipids. This study was designed to investigate the expression patterns, polymorphisms and associations with meat quality traits of the FABP1 gene in pigeons. 2. The temporal-spatial expression patterns showed FABP1 was widely expressed in all eleven tissues from 0–4 weeks of age, the expression level in the liver was the highest, followed by the small intestine and subcutaneous fat. 3. Five novel SNPs were found; all of them were synonymous and in Hardy-Weinberg equilibrium. Association analysis revealed that for the SNP of G161C, the AB and BB genotypes had higher (P ≤ 0.01) inosinic acid concentrations in breast muscle than the AA genotype. The BB genotype showed the highest (P < 0.01) intramuscular fat among the three genotypes, and significantly greater FABP1 mRNA levels were observed in the breast muscle of the BB genotype than in the AA and AB genotypes (P < 0.01). In the SNP C1376T, the AB and BB genotypes showed higher (P < 0.01) intramuscular fat than the AA genotype, and the relative mRNA expression level of the BB (P < 0.01) and AB (P < 0.05) genotypes was higher than that of the AA genotype in breast muscle. Correlation analysis implied that the FABP1 mRNA expression level was closely related to the inosinic acid (P < 0.05) and intramuscular fat content (P < 0.01). Oil red O staining of frozen sections of breast muscle on d 28 for SNPs G161C and C1376T also indicated that the BB genotype had the highest intramuscular fat content in both SNPs. In addition, correlation analysis implied the FABP1 mRNA expression level was closely related to inosinic acid (P < 0.05) and intramuscular fat content (P < 0.01). 4. The results suggested that FABP1 could be a potential candidate gene in marker-assisted selection for breeding pigeons with high-quality meat. 相似文献