Single nucleotide polymorphism analysis of the encoding region of EX-FABP gene and its association with fattiness trait in chicken

Fattiness is an important parameter to estimate meat quality, which has high heritability. In this experiment, F2 chickens derived from Broilers crossing to Silky were used to study the effect of extracellular fatty acid inding protein (EX-FABP) gene on abdominal fat accumulation. Five exons and partial introns of the gene were amplified by six pairs of primers, and then single nucleotide polymorphisms (SNPs) were detected by single strand conformation polymorphism (SSCP) and subsequently confirmed by sequencing. There were four nucleotides variations found, T-C at 659, C-T at 924, CA-TG at 968 and C-T at 1985 respectively. The result of least square analysis suggests that the chickens with HH genotype defined by the third pair of primers have a lower abdominal fat weight and abdominal fat percentage than the chickens with other genotypes (GG, GI, GH and II). It implies that extracellular fatty acid binding protein gene could be a candidate locus or linked to a major gene that significantly affects abdominal fat traits in chicken.     

猪生长激素cDNA基因的改建

为了将猪生称激素cDNA基因在表达后能一步纯化获得表达产物,对于来自质粒的猪生长激素cDNS基因通过PCR技术进步了改建,进一步的序列分析表明,与GenBank登录的序列不同,改建后的基因不带有突变。     

鸡体腹脂的遗传、选择方法和效果的研究进展

腹脂是鸡体蓄积脂肪的主要部位,肉用仔鸡的腹脂重和体重呈中等以上的表型相关和遗传相关,低向选择腹脂对肉用仔鸡的体重,生长速度及肉质不会产生明显影响。多项研究表明;血浆ＶＬＤＬ浓度对肉用仔鸡体腹脂进行间接选择是目前认为从遗传选择上控制鸡体脂肪沉只的既经济又现实可行的方法 。     

Fast evolution of growth hormone receptor in primates and ruminants

Pituitary growth hormone (GH) evolves very slowly in most of mammals, but the evolutionary rates appear to have increased markedly on two occasions during the evolution of primates and ruminants. To investigate the evolutionary pattern of growth hormone receptor (GHR), we sequenced the extracellular domain of GHR genes from four primate species. Our results suggested that GHR in mammal also shows an episodic evolutionary pattern, which is consistent with that observed in pituitary growth hormone. Further analysis suggested that this pattern of rapid evolution observed in primates and ruminants is likely the result of coevolution between pituitary growth hormone and its receptor.     

中国荷斯坦牛催乳素基因A8398G多态性与产奶性状相关性分析

采用PCR-RFLP技术对中国荷斯坦牛PRL基因第4外显子A8398G位点进行单核苷酸多态性分析。结果表明PRL基因经RsaⅠ酶切后产生2种等位基因和3种基因型。等位基因A和G的频率分别为0.257和0.743。基因型AA、AG和GG的频率分别为0.010,0.497及0.493。测序分析表明AA型和GG型相比在PRL基因第8398位碱基处发生A→G突变,该突变未导致氨基酸的改变。χ2适合性检验表明,PRL基因的RsaⅠ酶切位点处于Hardy-Weinberg不平衡状态(P<0.05)。在第Ⅱ泌乳期,最小二乘分析表明:基因型GG所对应的乳蛋白率最小二乘均值显著高于基因型AG所对应的最小二乘均值(P<0.05);基因型AG所对应的产奶量最小二乘均值显著高于基因型GG所对应的最小二乘均值(P<0.01);基因型GG所对应的线性评分最小二乘均值显著高于基因型AG所对应的最小二乘均值(P<0.05)。     

Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.     

Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.     

Rapid growth cost in ＂all-fish＂ growth hormone gene transsenic carp： Reduced critical swimming speed

Evidence has accumulated that there is a trade-off between benefits and costs associated with rapid growth. A trade-off between growth rates and critical swimming speed （Uc,t） had been also reported to be common in teleost fish. We hypothesize that growth acceleration in the F3 generation of ＂all-fish＂ growth hormone gene （GH） transgenic common carp （Cyprinus carpio L.） would reduce the swimming abilities. Growth and swimming performance between transgenic fish and non-transgenic controls were compared. The results showed that transgenic fish had a mean body weight 1.4--1.9-fold heavier, and a mean specific growth rate （SGR） value 6%-10% higher than the controls. Transgenic fish, however, had a mean absolute Ucr, （cm/s） value 22% or mean relative Ucrit （BL/s） value 24% lower than the controls. It suggested that fast-growing ＂all-fish＂ GH-transgenic carp were inferior swimmers. It is also supported that there was a trade-off between growth rates and swimming performance, i.e. faster-growing individuals had lower critical swimming speed.     

脊椎动物促性腺激素释放激素的结构、基因表达与调控的研究进展

综述了有关对脊椎动物促性腺激素释放激素的结构、基因表达与调控的研究进展 .阐明促性腺激素释放激素结构多样性 ,功能多样性 ,及其基因表达与调控 ,并对可能的应用前景作简要阐述     

胞内表达猪生长激素基因的重组酵母菌构建与鉴定

将猪生长激素(pGH)cDNA定向插入毕赤酵母胞内表达载体pPIC 3.5k质粒,重组构建了pPIC 3.5kpGH/GS115酵母表达系统, 其表达产物的SDSPAGE和Western blot结果表明, pGH蛋白在胞内获得正确表达,表达量达378 mg/L.     

Transgene for growth hormone in common carp（Cyprinus carpio L.） promotes thymus development

The transgenic carp were produced by micro-injection of CAgcGHc into the fertilized eggs. Observation of the thymus development between the transgenics and non- transgenic controls was carried out. The thymus of one-year- old transgenics F1 showed a great increase in both size and weight. The unilateral thymus of the transgenics weighed from 190 to 295 mg with average 218.6 mg, whereas the uni-lateral thymus of the controls weighed 20—81 mg with av-erage 42.5 mg; i.e. the thymus weight in the transgenics was 5.14 fold over that in the controls. The index of thymus/body weight in the transgenics was 2.97 fold over the controls. Light microscopy observation indicated that the thymus of the transgenics well developed with the thickened outer re-gion and compactly arranged thymocytes, while the thymus in the controls were degenerating with the thinned outer region, scattered thymocytes and groups of fatty cells. Fur-ther analysis with the electron microscopy revealed that pro-liferous cells in the transgenics were mainly small lympho-cytes and no pathological changes were found. The results confirmed that the 揂ll-fish?GH-transgene promotes thy-mus development and thymocyte proliferation, and retards thymus degeneration. The study has laid a foundation for further analysis of the immunobiological function in GH- transgenic carp.     

大黄鱼生长激素基因的克隆与表达

采用RT-PCR方法从大黄鱼脑垂体总RNA中克隆编码生长激素基因序列,并与数据库中鱼类生长激素基因进行比较.扩增获得的生长激素基因定向克隆到表达质粒pET-22b(+),并在大肠杆菌Rosetta(DE3)中表达.表达的蛋白为可溶性蛋白,表达量占细胞总蛋白的5%.     

Differential expression analysis and regulatory network reconstruction for genes associated with muscle growth and adipose deposition in obese and lean pigs

During the growth and development of skeletal muscle cells and adipose cells, the regulatory mechanism of micro-effect polygenes determines porcine meat quality, carcass characteristics and other relative quantitative traits. Obese and lean type pig breeds show obvious differences in muscle growth and adipose deposition; however, the molecular mechanism underlying this phenotypic variation remains unknown. We used pathway-focused oligo microarray studies to examine the expression changes of 140 genes associated with muscle growth and adipose deposition in longissimus dorsi muscle at six growth stages (birth, 1, 2, 3, 4 and 5 months) of Landrace (a leaner, Western breed) and Taihu pigs (a fatty, indigenous, Chinese breed). Variance analysis (ANOVA) revealed that differences in the expression of 18 genes in Landrace pigs and 3 genes in Taihu pigs were very significant (FDR adjusted permutation, P < 0.01) and differences for 22 genes in Landrace pigs and 7 genes in Taihu pigs were significant (FDR adjusted permutation, P < 0.05) among six growth stages. Clustering analysis revealed a high level of significance (FDR adjusted, P < 0.01) for 4 gene expression patterns, in which genes that strongly up-regulated were mainly associated with the positive regulation of myofiber formation and fatty acid biogenesis and genes that strongly down-regulated were mainly associated with the inhibition of cell proliferation and positive regulation of fatty acid β-oxidation. Based on a dynamic Bayesian network (DBN) model, gene regulatory networks (GRNs) were reconstructed from time-series data for each pig breed. These two GRNs initially revealed the distinct differences in physiological and biochemical aspects of muscle growth and adipose deposition between the two pig breeds; from these results, some potential key genes could been identified. Quantitative, real-time RT-PCR (QRT- PCR) was used to verify the microarray data for five modulated genes, and a good correlation between the two measures of expression was observed for both two pig breeds at different growth stages (r=0.876 ± 0.095). These results highlight some possible candidate genes for porcine meat quality and carcass traits and provide some data on which gene (s) should be further studied for elucidating the molecular mechanism of muscle growth and fat deposition.     

牛蛙生长激素基因cDNA克隆及在大肠杆菌中的表达

以成体牛蛙脑垂体总RNA为模板进行RT-PCR,克隆到牛蛙生长激素基因(bullfrog growth hormone, bfGH)cDNA编码序列,其长度为651 bp,编码的前体GH蛋白序列经BLAST分析,与已报道的牛蛙GH蛋白质序列AAB24792、AAB19428、CAA31038的同源性分别为98.1%、96.3%、95.3%,其在Genbank的登录号为AY251538;将bfGH亚克隆到原核表达载体pGEX1-λt构建成牛蛙GH原核表达载体VGBfGH,转化BL21（DE3）E.coli得     

云岭牛HMGA2基因InDel多态及其与生长性状的关联分析

为了探索高迁移率蛋白2(high mobility group AT-hook 2,HMGA2)基因插入缺失(InDel)多态与云岭牛体尺性状的关联性,以356头云岭牛为实验对象,采用多聚酶链式反应(polymerase chain reaction,PCR)扩增、聚丙烯酰胺凝胶电泳和Sanger测序的方法检测HMGA2基因的遗传变异,并利用SPSS软件对其InDel多态性与云岭牛体尺性状进行了关联分析.结果发现,在HMGA2基因的ARS-UCD1.2 5:47872723位点存在15 bp缺失,并根据缺失的类型,分为野生型(II)、杂合型(ID)、突变型(DD)(I:insertion,D:deletion),这3种不同的基因型所对应的个体,在腹围性状方面存在显著差异,而在其他的体尺性状上则不存在显著差异.因此,可以把HMGA2基因此位点InDel多态作为云岭牛腹围性状选择的候选分子标记,应用于肉牛新品种的标记辅助培育.     

柞蚕核型多角体病毒fgf基因的克隆和分析

为获得柞蚕NPV的全基因组序列,在柞蚕尸体中分离纯化柞蚕NPV,提取其基因组DNA,分别建立ApNPV DNA的HindⅢ和SalⅠ基因文库．对插入片段进行测序,经过同源性分析发现一个与人源fgf基因相似的基因,其读码框含有185aa．核苷酸和氨基酸同源性比较的结果表明：在genebank中没有同源性极高的序列,是个新基因;柞蚕NPV的fgf基因与云杉卷叶蛾NPV的同源性较高,分别为56．1％和45．9％,而与家蚕NPV及AcNPV的同源性相对较低,说明柞蚕NPV与家蚕NPV在进化上亲缘关系较远．该基因的克隆为进一步研究其在病毒感染过程中的作用打下基础．     

Therapeutic induction of angiogenesis by direct myocardial administration of an adenovirus vector encoding human hepatocyte growth factor gene and its safety

After the study in vitro and in rats, we assessed further the effects and safety of local angiogen therapy using intramyocardial delivery of an adenovirus carrying hepatocyte growth factor gene (Ad-HGF) in a canine ischemia model. The angiogenic activity of Ad-HGF was evaluated from three aspects. First, the augmentation of collateral vessel development was assessed by angiography 30 d after surgery. The results showed that the density of collateral vessels in treated group was higher than that of control group. Secondly, infarct size was evaluated by TTC staining and image analysis. The results showed that the infarct size of treated group was smaller than that of control group. Thirdly, the myocardial regional blood flow was determined by the method of colored microspheres. The results showed that the blood flow recovered to the level before ligation in treated group, but that of the control group was lower than normal level. In addition, during the study of chronic toxicity, we tested the anti-adenovirus antibodies by neutralization method. The antibodies yielded after the fourth injection decreased slowly from peak level and disappeared 12 weeks after drug withdrawal. Overall, Ad-HGF can promote angiogenesis in ischemic myocardium and reduce infarct size. So this method may be considered as a therapeutic angiogenesis induction strategy for ischemic disease including myocardial infarction and peripheral artery disease. At the same time, Ad-HGF could induce the yield of anti-adenovirus antibodies to neutralize adenovirus, which may be the mechanism of adenovirus clearance.     

Therapeutic angiogenesis induced by human hepatocyte growth factor gene in hindlimb ischemia of dogs

A preclinical study of treating peripheral srtery occlusive disease(PAD) was performed by using a hepatocyte growth factor(HGF) gene-expressing vector,plasmid pUDKH,in a dog model with complete ischemia of one hindlimb.After ligation of femoral artery of one hindlimb,pUDKH was transferred directly into the ischemic limb muscles.The angiogenic activity of the plasmid pUDKH was evaluated.On D 30 after injecting once of pUDKH at differ-ent doses into local muscles immediately after operation,the degree of augmentation of collateral vessel formation was significantly greater than that treated by blank vector.In addition,the blood flow rate of femoral artery in dogs treated with pUDKH was recovered on D90,while the folw rate was only 1/5 tp 1/3 in control dogs.The pulse amplitude of pUDKH-treated dogs was recovered on D90,but it was hardly detectable in most of the control dogs.The side effects of intramuscular transfection of pUDKH were also investi-gated,and no significant positive change was found.It is suggested that angiogenesis induced by HGF gene has the potential for clincal use in the treatment of peripheral arte-rial diseases.     

Mutations analysis of STK_(11) gene in Chinese families with Peutz-Jeghers syndrome

Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease that is characterized by multiple gastrointestinal hamartomatous polyps and melanin spots on lips and buccal mucosa at young age[1,2]. Previous studies have demonstrated that PJS predisposes carriers to cancers of gastrointestinal tract, uterus, ovary, testis, breast and other extragastrointestinal organs[3—5]. The STK11 gene, encoding a serine/threonine kinase at chro-mosome 19p13.3, was identified in 1998 as the main causativ…