VLDL双向选择肉鸡群SSR指纹分析

对初步进行VLDL双向选择的三个世代高、低脂群肉鸡进行SSR指纹分析,评定各基因座基因频率的变化,进而判断SSR标记与肉鸡肥度性状VLDL的相关关系,为低脂肉鸡的早期选育奠定基础。5个微卫星引物一个未扩增出产物,其余4个引物扩增出14个微卫星位点。各位点在高、低脂系中的基因频率经卡方检验,一世代有一个基因座的基因频率差异显著（P<0.05）;二世代两个基因座差异显著;三世代共检测到4个基因座差异显著。 Abstract:SSR fingerprints were analyzed in three generations of fat line (FL) and lean line (LL) of broiler chickens.Changes in gene frequencies of every locus were evaluated.Thus the relationship between SSR markers and VLDL (a trait representing fat mass of broiler),which is the basis for early selection of LL broiler,was examined.Fourteen microsatellite locus were successfully amplified with 4 of 5 primers used.The results of χ2 test for the gene frequencies of every locus show that one locus was significantly different in generation 1(P<0.05),two in generation 2 and 4 in generation 3.     

QTL Scanning for Rice Yield Using a Whole Genome SNP Array

High-throughput SNP genotyping is widely used for plant genetic studies. Recently, a RICE6K SNP array has been developed based on the Illumina Bead Array platform and Infinium SNP assay technology for genome-wide evaluation of allelic variations and breeding applications. In this study, the RICE6K SNP array was used to genotype a recombinant inbred line （RIL） population derived from the cross between the indica variety, Zhenshan 97, and the japonica variety, Xizang 2. A total of 3324 SNP markers of high quality were identified and were grouped into 1495 recombination bins in the RIL population. A high-density linkage map, consisting of the 1495 bins, was developed, covering 1591.2 cM and with average length ofl.1 cM per bin. Segregation distortions were observed in 24 regions of the 11 chromosomes in the RILs. One half of the distorted regions contained fertility genes that had been previously reported. A total of 23 QTLs were identified for yield. Seven QTLs were firstly detected in this study. The positive alleles from about half of the identified QTLs came from Zhenshan 97 and they had lower phenotypic values than Xizang 2. This indicated that favorable alleles for breeding were dispersed in both parents and pyramiding favorable alleles could develop elite lines. The size of the mapping population for QTL analysis using high throughput SNP genotyping platform is also discussed.     

猪Mu阿片受体基因单核苷酸多态性分析

Mu阿片受体（简称MOR）属于G蛋白藕联受体,分布在痛觉传导区,以及与情绪和行为有关的区域,影响动物的神经反应和行为表现。该研究以长白猪、大白猪和杜洛克猪为试验材料, 用8对引物对Mu阿片受体基因的5′ UTR区域、整个编码区和3′ UTR区域用PCR-SSCP方法进行了扫描,发现5处突变基因座（GenBank登录号：AF521309）。统计结果发现基因型频率分布与品种有关,大白猪突变基因型频率显著高于长白和杜洛克,本研究推测分布上的差异可能是由于长期的选择压力造成的。 Abstract:Mu opioid receptor (MOR) is a member of G protein-coupled receptor family,distributed in the pain transduction region in the brain and related to emotion and behaviour.This study was designed to investigate the Single Nucleotide Polymorphism (SNP) of Mu opioid receptor gene in various breeds,including duroc,landrace and Yorkshire.5′ UTR ( untranslate region),coding region and 3′ UTR of Mu opioid receptor gene were amplified by eight pairs of primers,and the Single Nucleotide Polymorphism (SNP) were detected by SSCP.Five polymorphisms were found (Genebank Accession number:AF521309).The results of χ2 test showed that the frequencies of genotypes in different breeds were significantly different (P＜0.01).The frequencies of mutation genotypes in Yorkshire were significantly higher than Duroc and Landrace.According to the above results,we can speculate the difference of the frequencies of genotypes may be the results of long term choice pressure.     

ZmcrtRB3 Encodes a Carotenoid Hydroxylase that Affects the Accumulation of α-carotene in Maize Kernel

α-carotene is one of the important components of pro-vitamin A,which is able to be converted into vitamin A in the human body.One maize(Zea mays L.) ortholog of carotenoid hydroxylases in Arabidopsis thaliana,ZmcrtRB3,was cloned and its role in carotenoid hydrolyzations was addressed.ZmcrtRB3 was mapped in a quantitative trait locus(QTL) cluster for carotenoid-related traits on chromosome 2(bin 2.03) in a recombinant inbred line(RIL) population derived from By804 and B73.Candidate-gene association analysis identified 18 polymorphic sites in ZmcrtRB3 significantly associated with one or more carotenoid-related traits in 126 diverse yellow maize inbred lines.These results indicate that the enzyme ZmcrtRB3 plays a role in hydrolyzing both α-and β-carotenes,while polymorphisms in ZmcrtRB3 contributed more variation in α-carotene than that in β-carotene.Two single nucleotide polymorphisms(SNPs),SNP1343 in 5 untranslated region and SNP2172 in the second intron,consistently had effects on α-carotene content and composition with explained phenotypic variations ranging from 8.7% to 34.8%.There was 1.7-to 3.7-fold change between the inferior and superior haplotype for α-carotene content and composition.Thus,SNP1343 and SNP2172 are potential polymorphic sites to develop functional markers for applying marker-assisted selection in the improvement of pro-vitamin A carotenoids in maize kernels.     

云南省三个地方鸡种血液淀粉酶多态性分析

采用垂直平板聚丙烯酰胺凝胶电泳法,测定了版纳斗鸡、武定鸡、尼西鸡的血液淀粉酶(Amy)。发现云南3个地方鸡种Amy-1的AA、BB型纯合子基因型频率很低, 其中尼西鸡为0,而AB型杂合子基因型频率很高,杂合子显著过量,均偏离Hardy-Weinberg平衡。在版纳斗鸡少数个体中还发现控制血液淀粉酶的另一位点Amy-2。 Abstract:Blood amylase polymorphism in 3 Yunnan local chicken breeds(Banna ganme chicken,Wuding chicken.Nixi chicken)was determined with polyacrylamide gel electrophoresis,It was found that frequen cies of both AA and BB homozygous genotype were very low,while AB heterozyote genotype frequency was very high in 3 Yunnan local chicken breeds and that Amy-1 distribution in these breeds was widely deviated from Hardy-Weinberg equilibrium proportion.We have also found that there is another locus Amy-2 which controls the amylase in a few Banna game chickens.     

中国地方鸡种血液淀粉酶Amy-1多态性

本文用聚丙烯酰胺水平凝胶电泳法测定了我国 10个地方鸡种的血液淀粉酶Amy-1多态性,发现我国地方鸡种Amy-1的AA、BB型纯合子基因型频率很低, AB型基因型频率很高,杂合子显著过量,且普遍存在Amy-1分布不平衡。还发现Amy-1A和Amy-1B两个等位基因频率在所有10个鸡种中几乎均为0.5。 Abstract:Blood amylase-1 polymorphism in 10 Chinese native chicken breeds was determined with polyacrylamide horizontal gel electrophoresis.It was found that both AA and BB genotype frequencies were very low,but AB genotype frequency was very high in Chinese native chicken breeds and that amylase-1 distribution in these breeds was widely deviated from Hardy-Weinberg equilibrum proportion.Both Amy-1A and Amy-1B gene frequencies were nearly 0.5 in all 10 breeds.     

不同猪种肠毒素大肠杆菌（ETEC）F4受体微卫星标记遗传差异性研究The

采用13号染色体上与K88ab和K88ac受体基因连锁的2对引物（S0223和S0068）,研究沙子岭猪和大约克猪的遗传差异性。结果表明,2个猪种在2个基因座均存在多态性,其基因杂合度和Shannon信息指数存在很大差异,而中外猪种的K88ab和K88ac受体基因也存在遗传差异,这2对引物可望作为K88ab和K88ac受体基因的遗传标记。Abstract: The genetic variation of ETEC F4 receptor in Shaziling and Yorkshire breeds were studied using two microsatellite markers(S0223 and S0068). The results showed that there were polymorphisms in the two markers, and there were great variations of the gene heterozygosity and Shannon information index in the two breeds. It was also reported that there were differences in K88ab and K88ac receptors in Chinese native breeds and foreign breeds, so the two markers might be the genetic markers of F4 receptor gene.     

Construction of a recombinant human GM-CSF/MCAF fusion protein and study on its in vitro and in vivo antitumor effects

A novel cytokine fusion protein was constructed by fusing granulocyte macrophage colony stimulat-ing factor (GM-CSF) with monocyte chemotactic activating factor (MCAF), which acts as a factor directing effector cells (monocytes) to a target site. The recombinant human GM-CSF/MCAF fusion protein could sustain the growth of GM-CSF-dependent cell line TF1 and was chemotactic for monocytes. The in vitro antitumor effect showed that rhGM-CSF/MCAF could activate monocytes to inhibit the growth of several human tumor cell lines, including a promyelocyte leukemia cell line HL-60, a lung adenocarcinoma cell line A549, a hepatoma cell line SMMC-7721 and a melanoma cell line Bowes. Furthermore, the cytotoxicity of monocytes activated by rhGM-CSF/MCAF against HL-60 and A549 was greater than that activated by GM-CSF or MCAF alone, even greater than that activated by a combina-tion of GM-CSF and MCAF, suggesting that the fusion protein has synergistic or enhanced effects. The in vivo anti-tumor effect indicated that     

Characterization and application of PBA fiber optic chemical film sensor based on fluorescence multiple quenching

The three types of structure of the pyrenebutyric acid of fiber optic chemical film sensor were stud-ied by fluorescence multiple quenching. They are, for different test samples and purposes, respectively general, three-way and combined. A tri-cup method was designed to demonstrate the multiple quenching of response mechanism, and a relationship formula of mathematical approach was established. The response mechanism was shown to include the dynamic quenching , inner-filter effects and/or resonance energy transfer. To show the response characterization in a series of organic and inorganic quenchers, a new concept of apparent quenching coefficient Kq was advanced. This kind of sensor has been used in continuous and in situ monitoring of the dissolution rate of drug tablets, on line and in situ monitoring of some organic therapeutic drugs in biological fluid and Cr( VI ) in industrial waste water. The measured data were examined and compared with HPLC or HPTLCS. Test results show that the sensors and appa     

Cloning and Characterization of the Microspore Development-Related Gene BcMF2 in Chinese Cabbage Pak-Choi （Brassica campestris L. ssp. chinensis Makino）

For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages, as well as in rosette leaves, florescence leaves, and scapes was analyzed using cDNA amplified fragment length polymorphism (cDNA-AFLP) in the genic male sterile A and fertile B line of Chinese cabbage pak-choi. Following amplification of 125 pairs of primer combinations, 11 differential fragments were obtained, of which eight were from the B line and the other three were from the A line. Of 11 differential fragments, four were verified by Northern hybridization that were expressed preferentially in fertile flower buds. Results of GenBank BLAST showed that one fragment was with unknown function, whereas the other fragments have strong nucleotide sequence similarities with the polygalacturonase (PG) gene, the pectinesterase (PE) gene, and the polygalacturonase inhibitory protein (PGIP4) gene. Only fulllength cDNA from the differential fragment BcMF-A 18T 16-1 was amplified by rapid amplification of cDNA ends (RACE) and Northern analysis showed that this fragment was expressed only in medium and largesized flower buds of the B line. The full-length cDNA, designated as BcMF2 (Brassica campestris Male Fertile 2), was 1 485 bp long and was composed ofa 1 263-bp open reading frame, which had 83% nucleotide similarity to a PG gene from Arabidopsis encoding polygalacturonase. Analysis of the basic structure of the protein revealed that it had one polygalacturonase active site (RVTCGPGHGLSVGS) at 256th site of amino acids and was classified as being a member of family 28 of the glycosyl hydrolases. The role of the BcMF2gene on microspore development is discussed in the present paper.     

鸭生长激素基因内含子2、3多态性分析

根据鸭生长激素基因内含子2、3的序列设计5对引物,利用PCR-SSCP方法对北京鸭、西湖野鸭、金定鸭、山麻鸭、荆江鸭、绍兴鸭等6个鸭品种进行了单核苷酸多态性分析, 并检测其多态性。结果共发现8个突变位点, 其中内含子2有7个: 2593处C-T, 2770处G-A, 2813处T-A, 2829处C-A, 2894处C-T, 2896处T-C,3100处C-G; 内含子3有1个: 3270处A-G。统计结果显示, 这8个变异位点的基因型频率分布与品种有关, 在这些基因座的变异水平上, 北京鸭和绍兴鸭表现出了相当的品种保守性, 本研究所检测到的这些基因座可能与鸭的生产性能有关。     

SNP variation in ADRB3 gene reflects the breed difference of sheep populations

The β3-adrenergic receptor (ADRB3), a G-protein coupled receptor, plays a major role in energy metabolism and regulation of lipolysis and homeostasis. We detect the single nucleotide polymorphism (SNP) variation in full-length sequence of ovine ADRB3 gene in 12 domestic sheep populations within four types by polymerase chain reaction-single strand conformation polymorphism and sequencing to reveal the breed difference. Twenty-two SNPs, 12 of which in the exon 1 and ten in the intron, were detected, and 12 new exonic and four new intronic SNPs were found. Most SNPs presented in Shanxi Dam Line and least ones in Dorset. The average SNP number in both meat and dual purpose for meat and wool breeds was significantly higher than general and dual purpose breeds for wool and meat. Frequency of each SNP in studied breeds or types was different. The 18C Del and 1617T Ins majorly existed in dual purpose breeds for wool and meat. The 25A Del, 119C>G and 130C>T were mostly found in the meat and dual purpose for meat and wool breeds. The 1764C>A more frequently presented in meat than in other types. The majority of variations came from within the populations as suggested by analysis of molecular variance. Close relationship presented among the Chinese and western breeds, respectively. In conclusion, SNPs of ovine ADRB3 gene can reflect the breed difference and within- and between-population variations, and to a great extent, the breed relationship.     

鸡瘦蛋白受体(OBR)基因内含子8单核苷酸多态性与体脂性状的相关研究

以东北农业大学高低脂双向选择系的第 6世代肉鸡为材料 ,鸡 7周龄时测定体重和腹脂重等屠体性状。根据鸡瘦蛋白受体基因内含子 8的序列 (GenBank登陆号 :AF2 2 2 783 )设计引物 ,用直接测序的方法检测多态性位点 ,用PCR SSCP的方法进行基因型分析 ,建立适合的统计模型对多态性位点产生的基因型与生长和体组成性状进行相关分析。结果表明 ,在第 50 0和 659位碱基同时发生了T—C、G—A突变。经最小二乘分析 ,3种基因型在腹脂重和腹脂率上差异显著 (P <0 0 5) ,BB型个体腹脂重和腹脂率显著高于AB型 (P <0 0 5) ,极显著地高于AA型个体 (P <0 0 1) ;3种基因型在肝重上差异显著 (P <0 0 5) ,且AA基因型个体的肝重显著低于AB和BB基因型个体。初步推断OBR基因可能是影响鸡脂肪性状的主效基因或与主效基因连锁 ,推测可以利用这个多态位点对鸡的体脂性状进行标记辅助选择     

Metabolism of very low density lipoproteins in genetically lean or fat lines of chicken

Metabolism of very low density lipoproteins (VLDL) has been compared in fat (FL) and lean (LL) lines of chicken. When refed after fasting, plasma triglyceride concentration reached a significantly higher plateau in FL, although their feed consumption was lower than in LL. Newly synthesized VLDL were studied using anti-lipoprotein lipase antibodies. VLDL triglyceride (TG) concentrations were increased by antibody injection and reached a higher concentration in FL plasma than in LL. Newly synthesized VLDL exhibited a similar lipid composition. Fatty acid profiles were also similar when birds ingested a very low fat diet. Comparison of in vitro affinity of lipoprotein lipase and VLDL from both genotypes did not reveal any difference in Km and Vmax. [14C]labelled VLDL from fat or lean donors were prepared and were injected into chickens from both genotypes. Fractional rate constants did not differ between lines. However, as plasma VLDL-TG pools were very different, plasma turnover was higher in FL than in LL. About 3-fold more VLDL-TG were incorporated in abdominal fat of FL than in LL. Difference in fattening between both genotypes seem to be due to both increased VLDL secretion and VLDL removal from plasma without difference in VLDL characteristics.     

Struggle to survive: aphid—plant relationships under low-light stress. A case of <Emphasis Type="Italic">Acyrthosiphon pisum</Emphasis> (Harris) and <Emphasis Type="Italic">Pisum sativum</Emphasis> L.

Light is primary source of energy and also plays signaling and regulatory roles in developmental processes and defense responses of plants. The aim of the study was to determine the performance, settling preferences, probing, and feeding behavior of Acyrthosiphon pisum on Pisum sativum grown in complete darkness (NL), with light at minimum level required for photoperiodic reaction (LL) and under full-light (FL) conditions. The effect of A. pisum infestation on metabolic status and defense responses of peas under FL, LL, and NL conditions was also determined. The population growth rate was limited on LL and NL pea plants as compared to FL plants. The reproductive period of aphids on LL and NL plants was eight times shorter than on plants growing in FL. In contrast to aphids on FL plants, the majority of A. pisum rejected LL and NL plants during settling. Aphid probing activities were not impeded on LL and NL plants but the probes were significantly shorter than on FL plants and consisted mainly of non-phloem activities. The analysis of tolerance of P. sativum to A. pisum showed that on FL plants, the number of aphids was nearly five times higher than on plants growing in low light (LL) at the end of the 2-week experiment but the tolerance index of FL plants was higher than that of LL plants. The contents of chlorophyll a, chlorophyll b, carotenoids, saccharides, and phenolics and the activity of β-d-glucosidase were notably lower in LL and NL plants than in FL plants. The increase in light intensity from complete darkness to the minimum level required for photoperiodic reaction did not stimulate evident changes in the measured plant biochemical parameters. These trends occurred in aphid-free (AF) and aphid-infested (AI) plants. However, under FL conditions, β-d-glucosidase activity and the content of saccharides were lower in AI plants than in AF plants. No differences in the measured plant biochemical parameters between AI and AF plants occurred under LL and NL conditions. The low β-d-glucosidase activity and low content of phenolics in the light-deprived plants that have reduced ability to photosynthesize show that under the biotic stress of aphid infestation plants invest in supporting basic metabolism rather than in defense against herbivores.     

家鸡腺苷琥珀酸裂解酶基因多态性与肌苷酸含量的关联及其与红原鸡的亲缘关系

根据腺苷琥珀酸裂解酶(adenylosuccinate lyase, ADSL)基因外显子2的序列设计引物, 用PCR-SSCP的方法对隐性白羽鸡、丝羽乌骨鸡、白耳鸡、藏鸡以及红色原鸡两个亚种进行了单核苷酸多态性分析, 并检测到了多态性, 表现为3种基因型, 对两种纯合子进行直接测序, 结果发现3484位碱基处发生C→T突变。对3种基因型的肌肉肌苷酸含量的最小二乘分析结果显示TT型(突变型)个体的肌肉肌苷酸含量极显著地高于CT型、显著地高于CC型个体, CT型个体也稍高于CC型, 但差异不显著, 初步推测该位点可能与肌肉肌苷酸含量有关。根据该多态位点的基因频率, 基于Nei氏的遗传距离运用NJ聚类法构建系统发生树, 进行家鸡与原鸡的亲缘关系分析, 结果发现, 丝羽乌骨鸡与白耳鸡的亲缘关系最近, 藏鸡和中国红原鸡亚种的亲缘关系也较近, 中国地方家鸡品种与中国红原鸡亚种的亲缘关系较近,而与泰国红色原鸡的亲缘关系较远,隐性白羽鸡与原鸡亲缘关系最远, 初步得出中国家鸡有自己独自的血缘来源的结论。     

Association of a missense mutation in the bovine leptin gene with carcass fat content and leptin mRNA levels

Previously, we have shown that alleles of the BM1500 microsatellite, located 3.6 kb downstream of the leptin gene in cattle, were associated with carcass fat measures in a population of 154 unrelated beef bulls. Subsequently, a cytosine (C) to thymine (T) transition that encoded an amino acid change of an arginine to a cysteine was identified in exon 2 of the leptin gene. A PCR-RFLP was designed and allele frequencies in four beef breeds were correlated with levels of carcass fat. The T allele was associated with fatter carcasses and the C allele with leaner carcasses. The frequencies of the SNP alleles among breeds indicated that British breeds have a higher frequency of the T allele whereas the continental breeds have a higher occurrence of the C allele. A ribonuclease protection assay was developed to quantify leptin mRNA in a separate group of animals selected by genotype. Animals homozygous for thymine expressed higher levels of leptin mRNA. This may suggest that the T allele, which adds an extra cysteine to the protein, imparts a partial loss of biological function and hence could be the causative mutation.     

Genetic Variation and Association of Insulin-Like Growth Factor Binding Protein-3 with Performance in Swine

Genetic variation of insulin-like growth factor binding protein-3 was analyzed in 17 pig breeds (14 native Chinese and 3 European). Using PCR-single strand conformation polymorphism, we found a polymorphism in intron 2, and this SNP was the combined mutation of G897T-G903A-C911T. The Chinese breeds carried a higher TAT/TAT genotype frequency (over 50%), except for Bamei (22%), Yujiang Black (0.0%), and Erhualian (10.0%); the European breeds had a higher GGC/GGC genotype frequency (Large White 1.67%, Landrace 13.89%, Duroc 0.0%). The allelic frequency of TAT in Chinese breeds was over 50%, except for Yujiang Black (12.5%); the allelic frequency of GGC was over 50% in all European breeds. The effect of genotype on 43 performance traits was investigated in one population (Lantang × Landrace). Pigs with the TAT/TAT genotype had higher B-point and C-point back-fat thickness than pigs with the GGC/GGC genotype. The TAT/TAT pigs also scored higher in meat color than the GGC/GGC pigs. These results implied that IGFBP-3 may affect meat quality and carcass traits.