蚁科5个属蚂蚁的分子系统学研究(英文)

[Objective] The aim of this study was to explore the taxonomic status and genetic relationship of ants at molecular level.[Method]Applying cyt b gene as a molecular marker,molecular phylogenetic analysis of 14 ant species of 5 genera(Camponotus,Formica,Polyrhachis,Pheidole and Crematogaster)in Formicidae was conducted.Partial sequences of cyt b gene in 14 ant species were analyzed with software MEGA,Clustal X and PAUP,and phylogenetic trees were constructed by Neighbor-Joining method(NJ)and Maximum-Parsimony method(MP).[Result]NJ tree and MP tree showed that the 14 ant species could be clustered into 5 branches.[Conclusion]The results of molecular phylogenetic analysis coincided with the views of traditional morphological taxonomy.     

不同H9N2亚型鸭流感病毒NS1基因克隆和功能进化分析(英文)

[Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the specific RT-PCR method, some strains of H9 subtype waterfowl influenza virus were isolated from the 12 to 20 day-old muscovy duck flocks without any clinical symptoms in different areas of Guangdong Province. Four of these strains, including A/duck/ZQ/303/2007(H9N2) (A3 for short), A/Duck/FJ/301/2007(H9N2) (C1 for short), A/Duck/NH/306/2007(H9N2) (D6 for short), A/duck/SS/402/2007(H9N2) (E2 for short), and a strain named A/duck/ZC/2007(H9N2) (L1 for short) from a muscovy duck died of avian influenza virus (AIV), were used for NS1 gene cloning and sequencing. Subsequently, the obtained NS1 gene sequences were compared with other NS1 sequences registered in GenBank, and the phylogenetic analysis was also conducted. [Result] When compared with the H9N2 AIV NS1 sequences in GenBank, the NS1 genes of the four AIV strains A3, C1, D6 and E2 displayed homologies ranging from 99% to 100% at nucleotide level, and 95% to 100% at amino acid level; while the NS1 gene of L1 strain displayed homology ranging from 94% to 97% at nucleotide level, and 93% to 98% at amino acid level. The phylogenetic tree demonstrated that A3, C1, D6 and E2 were highly resemblant, and L1 was closest to AY66473 (chicken, 2003). By comparison with the NS1 gene sequences of L1, AF523514 (duck), AY664743 (chicken) and EF155262.1 (quail) using DNAstar, A3, C1, D6 and E2 presented nucleotide variations at site 21(R→Q), 70, 71(KE→EG), 86 (A→S), 124 (V→M) and 225 (S→N), and amino acid variations at site 21, 70, 71 and 86 in dsRNA-dependent protein kinase (PKR) binding domain of NS1 gene, which induced the evident variations of antigenic determinant and surface probability plot of NS1 protein. [Conclusion] This study suggested that the amino acid sequence variation in PKR binding domain of NS1 protein had something to do with the virus pathogenicity.     

Genetic Variations Among Liriomyza sativae Blanchard Populations Indicated by β-tubulin Gene Sequences

To analyze the genetic differentiation of the host- and geo-populations of Liriomyza sativae Blanchard, the β-tubulin gene of 5 host-populations and 6 geo-populations of L. sativae was sequenced. Subsequently, the sequences were analyzed by biosoftware DNAStar and MEGA, and the phylogenetic trees constructed. The results obtained by the two softwares were similar, that is, the sequences of β-tubulin gene were more than 98% homologous, among the host- and geo-populations of L. sativae, with only 8 variable sites, but no insertions and deletions were detected. It seems that differentiations in β-tubulin gene among the host- and geo-populations of L. sativae are related to the hobby to hosts and the geographical distributions, respectively.     

广西巴马小型猪脂联素受体1和受体2 cDNA的克隆及序列分析(英文)

[Objective] To clone and analyze the sequence of Adiponectin receptor 1 (AdipoR1) and receptor 2 (AdipoR2) cDNA of Guangxi Bama mini-pig. [Method] The Adiponectin receptors cDNAs were amplified by RT-PCR using skeletal muscle total RNA as template and then ligated into pMD18-T vector after purification. The recombinant pMD18-T vector was transformed into the E.coli DH5α for identification and sequencing. And the results were compared with the cDNA sequence from other species. [Result] The fragments, 1 128 bp and 1 161 bp in size, were amplified by RT-PCR and respectively consistent with the coding sequence of AdipoR1 gene and AdipoR2 gene. The homology analysis showed that the sequences of AdipoR1 gene and AdipoR2 gene were respectively 99.8% and 99.7% homologous to the sequence of domestic pig reported in GenBank with one base and three base missense mutations correspondingly. [Conclusion] The AdipoR1 gene and AdipoR2 gene were successfully amplified from Guangxi Bama mini-pig, laying the foundation for the further study of the biological function of AdipoR genes and the design of novel drugs with AdipoR as target.     

马MxA基因第12外显子多态性检测(英文)

[Objective] The study aimed to establish a fast and accurate method to detect the polymorphism of the 12th exon of equine MxA gene. [Method] The 12th exon of MxA gene was amplified by mismatch PCR and the products were analyzed by restriction fragment length polymorphism (RFLP) to determine the point mutation at the 1 790 nt of MxA cDNA. The sequence of the PCR products was also analyzed. [Result] There were three genotypes (AA, AB and BB) in the 12th exon of equine MxA gene; the 2 081 nt of MxA cDNA mutated from G to C, correspondingly changing the 562th amino acid of the coding region of MxA protein from tryptophan to cysteine; the specific sequence of the PCR products amplified by mismatch PCR-RFLP was consistent with the analysis results of RFLP. [Conclusion] The mismatch PCR-RFLP was an easy method with accurate results to detect the polymorphism of the 12th exon of equine MxA gene.     

马IGF-Ⅰ基因3′端侧翼区PCR-SSCP多态性分析(英文)

[Objective] To explore the polymorphism of the 3′ flank region of equine IGF-Ⅰ gene. [Method] The 3′ flank region sequences of IGF-Ⅰ gene were amplified from genomic DNA of 270 horses, which included 4 types of Mongolian horse, Sanhe horse and Thoroughbred, and then analyzed by PCR-SSCP. [Result] Three genotypes (AA, BB and AB) were detected by PCR-SSCP and the distribution of genotypes of all research objects except Xinihe horse and Baerhu horse were in line with the "Hardy-Weinberg Law". [Conclusion] There was a polymorphic locus in the 3′ flank region of IGF-Ⅰ gene, which might affect the equine growth and development mechanism. The study is of important theoretical and practical significance to improve the performance and to develop equine industry.     

The Finding and Phylogenetic Evolution Analysis of Bovine Piroplasms in the Rasǒn Area of North Korea

The objective of this study was to investigate the epidemiology of bovine Piroplasms infections in the Rasǒn area of North Korea.The survey was carried out by light microscopic examination of Giemsa-stained blood smears,PCR,and phylogenetic evolution analysis of 128 blood samples collected from the Rasǒn area.The results showed that the infection rates of the small and large parasites were about 2.5 and 1.5% on average,respectively,in all Theileria sergenti and Babesia ovatapositive blood smears by microscopic examination of blood smears.The detection rate of T.sergenti Giemsa-stained smears was 43.75%,while that with PCR was 67.97%.The detection rate of B.ovata Giemsa-stained smears was 49.21%,while that with PCR was 71.88%.The sequence and phylogenetic analysis of DNA showed 98.84% homology between the 18S rRNA gene sequences of T.sergenti isolates from North Korean and that of Yanbian state from China,indicating the closest genetic relationship between both of them.Moreover,100% homology was shown between the 18S rRNA gene sequence of B.ovata isolates from North Korea and the published sequence AY081192 of GenBank,indicating the closest genetic relationship between both of them.This survey confirmed that Ras n is the endemic area of T.sergenti and B.ovata in North Korea.     

Yeast-Derived β-1,3-Glucan Substrate Significantly Increased the Diversity of Methanogens During In vitro Fermentation of Porcine Colonic Digesta

The β-1,3-glucan from yeast has been extensively examined for its immuno-enhancing effects in animals.However,investigation on the relationship among β-glucan,gut microbiota and immune-modulating effects remains limited particularly in pigs.Considering the critical roles of gut methanogens in the microbial fermentation,energy metabolism and disease resistance,we investigated the phylogenetic diversity of methanogens from fermented cultures of porcine colonic digesta with（G） or without（N） yeast β-glucan based on sequences of the archaeal 16S rRNA gene.A total of 145 sequences in the G library were assigned into 8 operational taxonomic units（OTUs） with the majority of sequences（114/145） related to strains Methanobrevibacter millerae or Methanobrevibacter gottschalkii with high identities ranging from 97.9 to 98.6%,followed by 23 sequences to Methanobrevibacter ruminantium,2 sequences to Methanobrevibacter smithii and one sequence to Methanobrevibacter wolinii.The 142 sequences in the N library were assigned to 2 OTUs with most sequences（127/142） related to strains M.millerae or M.gottschalkii with sequence identities ranging from 97.9 to 98.5%,and 15 sequences related to M.gottschalkii with 97.9% identity.Shannon diversity index showed that the G library exhibited significantly higher archaeal diversity（P0.05） and Libshuff analysis indicated the differences in the community structure between the two libraries were significant（P0.0001）.In conclusion,the current study provides evidence that addition of yeast β-glucan significantly increased the diversity of methanogens in in vitro fermented porcine colonic digesta.     

马MxA基因第13外显子的多态性研究(英文)

[Objective] To investigate the polymorphism of the thirteenth exon of MxA gene in 4 species of horse. [Method] The thirteenth exon of MxA gene fragments were amplified from genomic DNA of Sanhe horse, Xinihe horse, Wushen horse and Baerhu horse with the primers designed according to the MxA sequence announced in GenBank; the polymorphism of MxA gene was detected by PCR-SSCP and the products were sequenced. [Result] The polymorphism of the thirteenth exon of MxA gene appeared only in Wushen horse, the 2 081 nt of which mutated from guanine (G) to adenine (A) and the corresponding amino acid of which changed from glutamate (Glu) to alanine (Ala). [Conclusion] The study provided a basis for exploring the antiviral effect of MxA protein.     

绿头鸭线粒体ND2基因全序列测定及其研究

［目的］分析绿头鸭的系统进化关系。［方法］用PCR产物直接测序的方法，测得4只绿头鸭线粒体ND2基因全序列，结合GenBank中已公布河鸭属野鸭ND2基因全序列，基于N-J法和MP法构建河鸭属野鸭系统进化树。［结果］4只绿头鸭ND2基因序列完全一致，其全长为1 041 bp，碱基A、G、T、C含量分别为28.91%、13.35%、20.75%和36.98%，A＋T含量约等于C＋G 绿头鸭ND2基因序列与斑嘴鸭完全相同，与其他野鸭的同源性较高。系统进化树结果表明，绿头鸭与斑嘴鸭关系密切，两者共享一个单倍型，它们与棕颈鸭、针尾鸭及绿翅鸭同属于进化枝A。［结论］ 家鸭可能由绿头鸭和斑嘴鸭驯化而来。     

绿头鸭线粒体ND2基因全序列测定及其研究

1材料与方法1．1材料野生绿头鸭咀样4份（雌雄各2份），采自吉林省吉林市。1．2总DNA提取采用酚氯仿抽提法提取总DNA，0．7％琼脂糖凝胶电泳检测，-20℃保存。     

高邮麻鸭白细胞介素-2基因的克隆与序列分析

[目的]对高邮麻鸭白细胞介素-2（CIL-2）基因进行克隆和序列分析。[方法]以高邮麻鸭外周血淋巴细胞提取的总RNA为模板,根据已发表的鸭IL-2基因cDNA序列设计并合成1对引物,应用两步RT-PCR技术扩增出IL-2基因的特异性片段。将扩增片段插入pGEM-T-easy载体,并进行测序分析和结果验证。[结果]阳性克隆所含插入片段的DNA序列全长为433bp,与预期大小一致,含有1个423bp大小的开放性阅读框,共编码141个氨基酸的前体蛋白,N端21个氨基酸形成信号肽,成熟蛋白为120个氨基酸,分子量约为13.66kD,含1个糖基化位点。该序列与绿头鸭、绍兴鸭、固始鸭相应序列的同源性均为99.8%,与广州白鸭的同源性为99.3%,存在少量核苷酸差异。[结论]高邮麻鸭IL-2编码区基因相对高度保守,为其用于临床疾病治疗提供了一定参考。     

不同物种TYRP2基因完整编码区生物信息学分析

应用生物信息学和比较基因组学方法,比较分析了人、毛猩猩、绵羊、牛、猪、马、犬、斑马鱼、大熊猫、小家鼠、褐家鼠、鸭嘴兽、非洲爪蟾、原鸡、绿头鸭、斑胸草雀、黑猩猩、恒河猕猴、狨、灰短尾负鼠和兔21个物种TYRP2基因完整编码区(CDS),并对其遗传多样性、分子系统发育、编码蛋白的氨基酸序列、信号肽、跨膜结构域、疏水性/亲水性、二级结构等进行了预测。结果表明,从21个物种的42条基因序列检测到741个多态位点,共生成26种单倍型。物种间TYRP2基因序列编码区存在较丰富的遗传多样性,物种内该基因序列编码区则存在较强的保守性。TYRP2基因密码子有较强的偏爱性。根据核苷酸歧异度和遗传分化系数对物种的亲缘关系进行分析,结果表明,人与黑猩猩亲缘关系最近,小家鼠和褐家鼠亲缘关系最近。TYRP2基因多肽存在信号肽,有跨膜结构域,表现为疏水性,二级结构的主要组成元件为无规卷曲和α螺旋。     

杂交绿头野鸭屠宰性能的主成分分析

利用主成分分析对以绿头野鸭为父本的不同杂交组合的屠宰性能进行了研究。结果表明,6个指标综合成了3个互不相关的主成分,累计贡献率达94.95%,完全反映了原指标所提供的信息。综合评定结果显示,含1/2绿头野鸭血统、1/4北京鸭血统和1/4樱桃谷鸭血统的LBLY组合的杂交效果最好。     

鲈形目线粒体DNA蛋白编码基因的适用性分析

[目的]对线粒体DNA 13种蛋白质编码基因的系统进化分析能力进行了评估。[方法]单个基因和基因拼接序列比较,综合考虑序列的信息量和邻位连接法构建系统进化树的置信度。[结果]按分类阶元不同将基因都分成不同的4等。在鲈形目的科间阶元,最好的为ND6和cox2 好的序列为ND5和ND4 ND4L、ND3和ATP8差 包括Cyt b、cox1在内的其余6种基因为中等 在属内种间阶元,最好的为ATP6和cox2 好的序列为ND2和cox1 ND6、ND3和ATP8差 包括Cyt b在内的其余6种基因为中等。另外,分析揭示序列长度的增加可以提高系统进化树的置信度,且属内物种间比较时序列长度的影响小于高级阶元。[结论]线粒体DNA蛋白质编码基因的信息分布具不均一性,需分类阶元选择最佳基因和组合。     

亚洲黑熊四川亚种线粒体NADH脱氢酶亚基5和亚基6基因的克隆与初步分析

[目的]克隆与分析亚洲黑熊四川亚种线粒体NADH的脱氢酶亚基5和亚基6基因（ND5和ND6）。[方法]根据已报道的部分熊科动物ND5和ND6基因序列的相关信息设计引物,运用PCR技术,首次从亚洲黑熊四川亚种的肌肉组织总DNA中成功克隆了ND5和ND6基因的编码序列,分析了其序列特征,并与已报道的9种熊科动物进行比较分析。[结果]该克隆片段包括ND5和ND6两个基因共2456bp,共编码781个氨基酸,与9种熊科动物的ND5和ND6具有很高的同源性;该克隆片段ND5和ND6的蛋白分子量分别为68.2621和19.0386kDa,等电点分别为9.19和4.25,与9种熊科动物的均十分接近。[结论]四川黑熊与9种熊科动物的ND5和ND6基因及其编码蛋白具有很高的相似性,在功能上具有一致性,尤其与东北黑熊在功能上具有高度一致性。     

线粒体ND5·Cytb部分序列在虎物种鉴定中的应用

[目的]根据线粒体ND5、Cytb基因部分序列,对送检的4件疑似虎的样品进行物种鉴定。[方法]利用PCR法,扩增样品的线粒体ND5、Cytb基因部分序列,对目的片段测序,通过blast比对,确定物种。[结果]4个样品的物种来源均为虎,但无法确认其亚种。[结论]虎亚种间的线粒体差异很小,并且存在不同亚种杂交的情况,只根据线粒体基因难以进行亚种鉴定。     

赤眼鳟线粒体NADH脱氢酶6基因克隆及序列特征分析

[目的]根据赤眼鳟线粒体NADH脱氢酶6（ND6）基因序列分析不同种鱼类的分子进化关系。[方法]根据其他鱼类的线粒体ND6基因及侧翼序列设计引物,采用PCR扩增并克隆、测序赤眼鳟ND6基因,通过MEGA4．0Neighbor—Joining法分析不同鱼类的进化关系。[结果]赤眼鳟（Squaliobarbus curriculus）线粒体基因组ND6基因序列长度为522bp,编码173个氨基酸。该基因碱基组成为：A41．5％、C32．7％、G12．4％、T13．4％,其中A＋T含量（54．9％）高于G＋C含量（45．1％）。将赤眼鳟ND6基因序列与GenBank中其他11种鱼类ND6基因序列进行比对发现,赤眼鳟ND6核酸序列与团头鲂的同源性最高,为90．8％;与红点鲑鱼的同源性最低,为73．0％。赤眼鳟ND6氨基酸序列与其他物种ND6氨基酸进行比对发现,赤眼鳟与团头鲂同源性最高,为98．3％;与大黄鱼的同源性最低,为75．7％。以ND6氨基酸序列采用NJ法构建12种鱼类的系统发生树基本反映了它们的种间、属间和科间的关系。[结论]首次得到了赤眼鳟线粒体ND6基因序列,并确定了赤眼鳟与其他11种鱼类的分子进化关系。     

保存温度与时间对动物组织DNA提取质量的影响

［目的］研究不同保存温度、不同保存时间对蟹肉总DNA提取质量的影响，为同类研究提供借鉴。［方法］将样品在4、-20和-40℃下分别保存4、8和15 d，提取螯足内肌肉总DNA，测定总DNA的紫外吸收值，并设计引物扩增其mtDNA ND6基因，采用琼脂糖凝胶电泳分别对提取的总DNA及PCR结果进行检测。［结果］由鲜活蟹组织提取的DNA A260与A280的比值因蛋白污染而较低；15 d内4 ℃与-40 ℃条件下保存的DNA提取率和条带检出率均无明显差异，而-20 ℃下的保存效果受时间影响明显。各处理组合总DNA的 A260与A280比值在1.79～2.00；利用上述DNA作模板，扩增其特定基因mtDNA ND6均可获得预期长度的PCR条带。［结论］短期保存组织宜选择4 ℃，若需较长时间保存时则宜选择-40 ℃以下的低温。