基因Ⅶ型新城疫病毒分离鉴定及免疫原性研究

旨在分离筛选免疫原性好的新城疫病毒(NDV)流行毒株。从发病鸡组织中分离到4株病毒,经血凝和血凝抑制试验鉴定为NDV,进而完成致病指数测定、F基因高变区测序和遗传进化分析,并通过鸡胚交叉中和试验和攻毒试验研究PLK-N-06株对比La Sota疫苗株的抗原差异和免疫保护效力。结果显示,4个分离株的致病指数均属速发型NDV范围,且PLK-N-06株毒力最强;F蛋白裂解位点均为112R-R-Q-K-R-F117,符合强毒株特征。遗传进化分析表明,分离株均为Ⅶd亚型。交叉中和试验结果显示,PLK-N-06株和La Sota株存在显著的抗原性差异。用PLK-N-06株油乳剂灭活苗免疫接种SPF鸡后产生的HI抗体效价几何平均值为1∶338,显著高于La Sota疫苗(1∶69),且其对PLK-N-06株感染的排毒率为0/10,明显低于La Sota株油乳剂灭活苗的4/10,结果表明PLK-N-06株具有良好的免疫原性,为新城疫基因Ⅶ型新型疫苗的研发奠定了基础。     

FABPs作为猪肌内脂肪沉积候选基因的研究进展

对FABPs 作为猪肌内脂肪沉积候选基因的研究现状及前景作一综述。主要包括H-FABP和A-FABP基因的性质、变异以及与肌内脂肪含量的关系。     

伪狂犬病病毒鄂A株TK基因的克隆及其鉴定

合成了 １ 对能对伪狂犬病病毒（ Ｐｓｅｕｄｏｒａｂｉｅｓ ｖｉｒｕｓ, Ｐ Ｒ Ｖ） Ｔ Ｋ（ｔｈｙｍ ｉｄｉｎｅ ｋｉｎａｓｅ）基因＋ １１９～＋ １ ０７１区进行特异扩增的引物,用猪 Ｐ Ｒ Ｖ 鄂 Ａ 株细胞培养物提取的基因组作模板,扩增出 ９５３ ｂｐ 长的片段,用地高辛标记该片段作探针,通过 Ｓｏｕｔｈ ｅｒｎ 杂交,从克隆有 Ｐ Ｒ Ｖ 鄂 Ａ 株的 Ｂａｍ Ｈ Ｉ片段的重组质粒中钓出含 Ｔ Ｋ 基因的重组质粒 ｐ Ｓ Ｔ Ｋ。对 ｐ Ｓ Ｔ Ｋ 进行酶切分析,绘制了含 Ｔ Ｋ 基因的 Ｂａｍ Ｈ Ｉ片段的图谱,通过测序得出了 Ｔ Ｋ 基因的全序列。将该序列与 Ｐ Ｒ Ｖ Ｎ Ｉ Ａ３ 株 Ｔ Ｋ 基因进行比较,发现鄂 Ａ 株的 Ｔ Ｋ 基因存在变异。     

口蹄疫病毒3ABC基因的克隆与测序

参考 Gen Bank中发表的猪源 O型口蹄疫病毒 3 ABC的基因序列 ,设计一对特异引物 ,以猪源 FMDV/ O/ CC株基因组 RNA为模板 ,RT-PCR扩增 3 ABC基因 ,并克隆到 p MD1 8-T载体中。测序结果显示 ,FMDV/ O/ CC株 3 ABC基因 c DNA长 1 2 81 bp,编码为 42 7个氨基酸残基组成的多肽。核苷酸序列和推导氨基酸序列同源性比较发现 ,FMDV/ O/ CC株和 FMDV/O/ TW/ 99株 3 ABC基因亲缘关系密切。核苷酸同源性为 97% ,推导氨基酸序列同源性为96.3 %。     

树鼩Tssk6基因cDNA克隆、组织表达谱及生物信息学分析

本研究旨在克隆出树鼩Tssk6基因cDNA的全长序列,阐明其组织表达谱及基本生物信息学特征。以GenBank中收录的树鼩Tssk6基因mRNA预测序列为参考设计特异性引物,对树鼩Tssk6基因的cDNA全长序列进行克隆,用实时荧光定量PCR检测该基因在树鼩不同组织中的表达情况,并对cDNA序列的CDS区核苷酸序列与蛋白质结构进行了生物信息学分析。结果显示,树鼩Tssk6基因cDNA的CDS区序列长度为822 bp,编码273个氨基酸;Tssk6基因仅表达于树鼩睾丸组织;对树鼩和其他8种哺乳动物Tssk6基因cDNA的CDS区核苷酸序列构建的系统进化树显示,树鼩与人、黑猩猩、恒河猴3种灵长类动物亲缘关系较近,与小鼠、大鼠、金仓鼠3种啮齿类动物亲缘关系较远。本研究为进一步研究树鼩Tssk6基因的功能奠定了基础。     

贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析

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。     

青海巨型住肉孢子虫线粒体cox1基因的克隆及进化分析

为了研究青海省巨型住肉孢子虫线粒体细胞色素C氧化酶第一亚基(cox1)基因部分序列(pcox1)的遗传变异情况,并用pcox1序列构建巨型住肉孢子虫与其他住肉孢子虫的种群遗传关系。利用PCR技术扩增巨型住肉孢子虫的pcox1,将测序所获序列进行比对,并进行同源性分析和系统发育树的构建。结果显示,所获得的pcox1序列长度均为968 bp,与GenBank已公布的巨型住肉孢子虫相似度为99.1%~99.8%。系统发育树表明,所有分离株与已知巨型住肉孢子虫位于同一分支。巨型住肉孢子虫pcox1序列种内相对保守(0.2%~0.7%),种间差异较大(6.8%~20.8%),可用作种间遗传变异研究的标记。     

Association Analysis Between Polymorphism of IGF1 Gene and Growth Traits in Rabbit

This study was aimed to analyze the association of insulin-like growth factor 1 (IGF1) gene polymorphisms with growth traits in rabbit. The technology of DNA pooling, PCR-SSCP and direct sequencing were used to detect the SNP of IGF1 gene. The results showed that T365C was detected in IGF1 gene exon 3 of rabbit. The SNP (C/T) was devised into three genotypes:TT, TC and CC. The relationships between genotypes and growth traits revealed that the birth weight and the weight at 28 days of age in New Zealand rabbit of TT genotype of T365C locus were significantly higher than TC and CC genotypes (P<0.05), the birth weight, the weight at 90 days of age and average daily gain form birth to 90 days of age in Belgian rabbit of TT genotype of T365C locus were significantly higher than TC and CC genotypes (P<0.05), there were no significant difference of the growth traits in other rabbit breeds (P>0.05).The consequence indicated that IGF1 gene was primarily deduced to be a potential major gene or linked to major gene effecting the growth traits of rabbit, and this SNP (T365C) might be a candidate molecular genetic markers to improve the growth traits of rabbit.     

Study on the Virulence Gene Distribution and Genetic Evolution of Cattle Shiga Toxin-producing Escherichia coli Isolates

This study was aimed to understand the relationship of virulence gene distribution and genetic evolution between cattle originated Shiga toxin-producing Escherichia coli (STEC) and human originated enterohaemorrhagic Escherichia coli (EHEC) O157. This experiment collected 18 strains STEC in a dairy farm from Jiangsu province and 9 STEC reference strains (human, sheep, swine and avian), according to the method of U.S. Centers for Disease Prevention and Control Center (PulseNet), using the XbaⅠ enzyme digestion and pulsed field gel electrophoresis (PFGE) analysis, virulence genes were detected in some STEC isolates. The virulence gene distribution of O157 from different origin was remarkably different. The cattle originated STEC O157 and the human originated EHEC O157:H7 (EDL933W) had the most similar virulence gene distribution. In contrast, virulence genes were lack in cattle STEC O18 and O26, even though the cattle STEC O18 and O26 had the similar genotype as human EHEC O157:H7 (EDL933W). PFGE of Xba Ⅰ digested chromosomal DNA from 27 isolates of STEC exhibited 22 profiles. In general,the Dice coefficients of different originated STEC ranged from 72% to 100%.Cattle STEC O157 had a high similarity with two strains of human originated EHEC O157, while a low similarity was demonstrated between cattle STEC O157 and STEC O157 of swine and avian. The Dice coefficients of the cattle STEC O157 and the two strains of human EHEC O157 ranged from 83% to 95%. The Dice coefficients of cattle STEC O26 (Ⅶ,Ⅷ) and the two strains of human EHEC O157 were more than 82%. Therefore, it was concluded that the cattle STEC O157 and human EHEC O157 had a closer relationship in terms of virulence gene distribution and in genetic evolution.