蛙皮素样肽家族及其受体氨基酸序列信息学比较分析

【目的】研究反刍动物蛙皮素样肽家族多肽及其受体的保守性。为不同动物,特别是反刍动物这2种蛙皮素样多肽及其受体蛋白抗原设计和活性多肽筛选等研究提供参考。【方法】采用生物信息分析学的方法,通过UniProt数据库比较牛的蛙皮素样肽家族胃泌素释放肽(Gastrin-releasing peptide, GRP)和神经介素B(Neuromedin B, NMB)2种多肽及其特异性受体GRP-R和NMB-R与其他动物氨基酸序列的差异性。【结果】13种动物成熟GRP多肽C-端的8个氨基酸序列完全相同,牛GRP氨基酸序列与绵羊、猪和豚鼠最高,分别为88.7%、77.8%和77.8%,与其他动物相似性低于74.1%。10种动物成熟NMB多肽C-端10个氨基酸序列完全相同。牛GRP-R与狗、马和猪等相似性最高,分别为96.1%、94.3%和94.0%,牛NMB-R与猪、人和马相似性最高,分别为92.3%、91.5%和91.0%;牛GRP-R与NMB-R相似性仅为62.2%。【结论】GRP和NMB的C-端氨基酸序列在不同动物之间均具有高度的保守性,而N-端为变化区域,且GRP-R和NMB-R氨基酸序列在动物之间保守性较高。     

肉牛源细极链格孢菌的分离鉴定

为鉴定肉牛皮肤病病原及其致病性并筛选其敏感药物,本研究取患皮肤病肉牛病变部位皮屑、毛发和痂皮进行病原菌的分离,并对分离菌株进行了形态学鉴定、ITS序列分析、小鼠致病性试验和药敏试验。结果显示分离到1株形态特征与细极链格孢菌极其相似的菌株(PY2-1-2);经PCR扩增、测序和BLAST序列比对,结果显示分离株(PY2-1-2)的ITS基因序列与细极链格孢菌(MG975630.1)的ITS基因序列的相似性为99%,分离株PY2-1-2的ITS基因序列长度为543 bp,该片段包括ITS1、5.8S rDNA和ITS2的全部基因序列以及18S rDNA和28S rDNA的部分基因序列,提交GenBank(MH656780.1)。根据形态学结合ITS基因序列分析结果确定该分离株为细极链格孢菌。小鼠致病性试验结果显示该菌对小鼠有致病性。药敏试验结果显示:该菌对灰黄霉素、氟康唑、伊曲康唑、酮康唑的最小抑菌浓度(MIC)值分别为0.5μg/mL、4μg/mL、1μg/mL、1μg/mL。本研究为今后进一步探究细极链格孢菌引起的皮肤病诊治提供科学有效的参考。     

一起山羊地方性鼻内肿瘤的诊断

陕西省某山羊养殖场发生一起山羊鼻内肿瘤病,通过流行病学和临床症状调查、病理剖检、病理切片检查进行初步诊断。其次根据GenBank收录的山羊地方性鼻内肿瘤病毒基因组序列设计1对特异性引物,通过PCR方法从肿瘤组织中扩增获得目的片段并测序。序列比对分析表明,该目的基因和山羊地方性鼻内肿瘤病毒高度相似,最终确定该病为山羊地方性鼻内肿瘤病毒引起的山羊地方性鼻内肿瘤(ENT)。     

柴达木盆地梭梭LEA基因的扩增及序列分析

为发掘梭梭(Haloxylon ammodendron)抗逆相关基因,以柴达木盆地梭梭的同化枝为材料,扩增其LEA基因并进行了序列测定与分析。结果表明,所得柴达木盆地梭梭的LEA基因碱基数为234 bp,其中A碱基70个,T碱基52个,G碱基59个,C碱基53个;氨基酸序列分析表明,所得LEA基因片段编码78个氨基酸,其中强碱性氨基酸有12个,强酸性氨基酸有6个,疏水性氨基酸有34个,亲水性氨基酸有26个。     

Molecular Cloning,Sequence Analysis and Tissues Expression Profile of Part of Mink Toll-like Receptors (TLRs) Genes

The present study was aimed to explore the potential role of Toll-like receptors (TLRs) on the mink immunity, and accumulate alternative genetic material for the mink breeding for disease-resistant.Four pairs of primers were designed according to ferret TLR sequences in GenBank to obtain the sequences of TLR genes (TLR4, TLR6, TLR7 and TLR8) of mink and then performed bioinformatics analysis of these sequences.In addition, the expression of TLR4 and TLR7 genes in various tissues in young and adult minks were analyzed by RT-PCR. Sequence homology analysis showed that mink had higher homology with carnivora (ferrets, polar bear, panda, walrus, seals, dog, tiger and cat) which had the nearest perimeter in the phylogenetic tree.Tissue expression analysis showed that TLR4 and TLR7 were widely and differently expressed in variety tissues of mink.Interestingly, the expression of TLR4 and TLR7 in young mink were much higher than that in adult mink.     

Cloning and Sequence Analysis of Complete Genome of Porcine Circovirus Type 2 from Liaoning Province

It was aimed to lay a foundation for epidemiological survey of porcine circovirus type 2(PCV2) in Liaoning province and selection of highly homologous strain inactivated vaccine.Some pigs suspected of having postweaning multisystemic wasting syndrome (PMWS) from Shenyang,Chaoyang,Tieling,Dalian and other places in Liaoning province,were detected by PCR method,then the whole genome of 10 positive samples were cloned and sequenced.The result showed that all of them were virulent strains,9 strains were 1767 bp and 1 strain was 1768 bp,and there was a T base additon at 1039 bp.The nucleotide homology was 95.1% to 98.5% with HM038034 and JQ692110.In the phylogenetic tree,JHZ2 was PCV2a type,the other 9 strains were PCV2b type,DLS38,LYD32 and SYX7 were in the same branch and same onset time.The amino acid homology of ORF2 was 89.6% to 100.0%,there were 35 mutation points,large degree of variation,the only glycosylation site (NYS) was conserved.10 strains did not change significantly,mutants were not due to PCVD,PCV2b was the predominant type,and PCV2 was related with seasonality.     

棉花单核苷酸多态性标记研究进展

单核苷酸多态性标记已在农作物研究中得到广泛应用并取得重大进展。为了便利棉花SNP(Single nucleotide polymorphism)标记的研究和应用,介绍了利用基因芯片、简化基因组测序、重测序等在棉花中开发SNP标记的方法 ,综述了SNP标记在棉花遗传图谱构建、数量位点的定位和分子标记辅助育种、基因组测序以及系统进化等研究中的应用。并对异源四倍体棉花中SNP标记开发时,同源序列位点和部分同源序列位点上的SNP标记辨别问题进行了系统探讨,对其快捷的开发、检测方式和在数量基因定位中的应用前景进行了展望。     

1株库蠓源版纳病毒的分离与全基因组序列分析

为分析云南省虫媒病毒的种类与遗传特征,在云南省师宗县采集库蠓进行病毒的分离与鉴定;通过全长cDNA扩增与高通量测序技术获取病毒全基因组序列,进行序列比对与系统发生树构建。结果显示,从采集的库蠓样本中分离出1株可在C6/36细胞上引起细胞病变的毒株（YNSZ043）,病毒基因组为分节段双链RNA,琼脂糖凝胶电泳呈"2-4-3"的带型特征;电镜观察可见直径为70~80 nm,呈"指环状",表面具有纤维突起的病毒粒子。全基因组测序结果显示,YNSZ043毒株为版纳病毒（Banna virus,BAV）,基因组大小为20 683 bp,由Seg-1（3 762 bp）至Seg-12（861 bp）12个基因节段组成,与中国BAV毒株各基因节段的核苷酸序列相似性在64.8%~99.6%之间,氨基酸序列相似性在58.8%~100%之间,在系统发生树上YNSZ043毒株与中国分离的BAV聚为一簇,形成独立的中国进化支系。对决定BAV基因型的Seg-12分析结果显示,YNSZ043毒株属于A2基因型,该毒株的Seg-5/VP5与越南分离BAV毒株的核苷酸和氨基酸序列相似性高达97.1%和97.6%,表明该毒株的Seg-5基因节段很可能与越南毒株之间发生了基因重配。研究结果丰富了中国BAV的基因组序列,为开展云南省BAV的流行病学研究提供了参考。     

Cloning and Sequence Analysis of HA and NA Genes of One Strain of H6N6 Subtype Avian Influenza Virus Isolated from Guizhou Province

To investigate the epidemic situation of H6N6 subtype avian influenza virus (AIV) in Guizhou province,A/duck/Guizhou/013/2014 was isolated from Sansui duck in live poultry market of Guizhou in 2014,the hemagglutinin (HA) and neuraminidase (NA) genes of DK/GZ/14 were subjected to clone and sequence analysis.The results showed that HA gene had the highest nucleotide homologies (97.5%) with the duck-origin H6N6 subtype AIV isolated from Eastern China in 2009,and the strains of HA gene proteolytic cleavage sites was P-Q-I-E-T-R-G,which accordeol with the molecular characteristic of low pathogenic AIV (LPAIV).However,NA gene of A/duck/Guizhou/013/2014 had the highest nucleotide homologies (98.2%) with the duck-origin H6N6 subtype AIV isolated from Fujian in 2007.The phylogenetic tree showed that A/duck/Guizhou/013/2014 and Hunan strains located in the same branch,while three duck-origin H6N6 subtype AIV isolated from Guizhou in 2007 and A/duck/Guizhou/013/2014 located in the different branch for HA and NA genes in genetic evolution,which suggested that A/duck/Guizhou/013/2014 was far with the local H6N6 subtype.The results also clearly indicated that duck-origin H6N6 subtype AIV had genetic diversity in duck population in Guizhou.