西藏蟾蜍线粒体COI和Cyt b基因序列的分析研究

采用聚合酶链式反应克隆西藏蟾蜍Bufo tibetanus线粒体COI和cyt b基因,首次报道该物种这两个基因的全序列,利用分子生物学软件结合比较与其他7种两栖动物的同源序列.结果显示:西藏蟾蜍两个基因序列中碱基G含量明显低于其它三种碱基,密码子第三位碱基G含量在4种蟾蜍中是最低的;碱基替换主要发生在第三位,属内转换率大于颠换率;相比核苷酸数据,氨基酸序列显示的遗传距离表明氨基酸序列更加保守,遗传距离显示西藏蟾蜍与中华大蟾蜍B.gargarizans的遗传距离最小;构建系统进化树,显示西藏蟾蜍和中华大蟾蜍的亲缘关系最近.     

中外六个猪种FUT1基因多态性研究

目的对6个中外猪种共245头猪的FUT1基因进行了研究。方法采用PCR-RFLP技术。结果与结论Hin6I位点上,大白猪、长白猪、杜洛克猪3个外来猪种均存在多态,且以敏感型（GG型和AG型）居多;山西黑猪、太原花猪、马身猪3个本地猪种的所有检测样品都表现为GG型。     

从Cyt b基因序列探讨鹿亚科动物的系统发生关系

鹿亚科共有4个属,各属间和属内特别是鹿属内的系统进化关系存在疑义。根据Cyt b基因序列分析,探讨了鹿亚科及中国鹿属、马鹿亚种的进化关系。结果分析表明：现行分类系统中,斑鹿属可能并非单系发生,暗示应将豚鹿并入鹿属;麋鹿属与鹿属有较近的进化关系,也应并入鹿属;黄占鹿属的进化地位有待进一步研究;归并后的鹿属为单系发生。中国马鹿各亚种在系统发生上是一单系群,其中马鹿天山亚种和阿尔泰亚种聚为最原始的一支。     

松江鲈鱼Cytb基因序列分析及种质鉴定

采用PCR方法扩增四尾松江鲈鱼线粒体细胞色素b(mitochondrial cytochrome b,Cytb)基因序列,全序列长度为1 141 bp,并建立了松江鲈鱼的实时定量PCR检测方法对该物种进行种质鉴定。分析Cytb基因序列基本特征显示,Cytb基因在第3位密码子表现出明显的反G偏倚,显示出脊椎动物Cytb基因的共同特性。第2位密码子嘧啶的含量远高于嘌呤的含量,有5个位点发生转换,0个位点发生颠换。选取杜父鱼科(Cottidae)的10种鱼类,与松江鲈鱼进行序列分析,构建发育系统树并分析遗传距离,与鲈鱼的亲缘关系最远。以松江鲈鱼Cytb基因序列为靶基因,利用PRIMER EXPRESS2.0软件设计引物和Taqman探针,建立检测松江鲈鱼的定量PCR方法,特异性强,灵敏度高,检测限为3.20×102拷贝/反应。     

中国缘蝽科部分种类细胞色素b基因序列及系统进化研究

以线粒体细胞色素b(Cyt b)基因作为分子标记,首次对缘蝽科4亚科14种昆虫进行序列测定,获得Cyt b基因412 bp的序列片段,该片段中碱基T、C、A、G的平均含量分别为34.2%、11.4%、35.7 %和18.7 %,A T平均含量为69.9 %,明显高于G C含量(30.1 %).密码子第3位点的A T含量高达82.8%,亚科间序列变异大,有193个核苷酸位点发生变异,碱基替换多发生于第3位点.以筛豆龟蝽为外群构建系统发育树,表明在亚科级关系上,姬缘蝽亚科最原始,蛛缘蝽亚科次之,巨缘蝽亚科和缘蝽亚科亲缘关系较近,为较进化种类.     

一枚恐龙蛋内细胞色素b基因片段的序列分析

用一对根据人线粒体细胞色素ｂ基因保守区序列设计的引物,以一枚恐龙蛋内ＤＮＡ为模板,在ＰＣＲ上扩增出大约１７０ｂｐ的基因片段。序列测定后经ＩＮＴＥＲＮＥＴ联网的ＢＬＡＳＴ数据库类似性检索,结果表明,该序列与人类线粒体细胞色素ｂ基因的序列完全相同（９６－１００％相似）。因此,可以肯定实验材料本身有严重污染。     

Association of Lymphotoxin Alpha Polymorphism with Type 1 Diabetes in a Tunisian Population

We investigated the association of the lymphotoxin (LT)-α gene polymorphism +249A/G with type 1 diabetes. The distribution of genotypes of the LT-α +249A/G single nucleotide polymorphism (SNP) was assessed in 115 diabetic patients and 123 normoglycemic control subjects, using PCR-restriction fragment length polymorphism analysis. Among unselected patients, the SNP was significantly associated with increased risk of diabetes (χ² = 8.44, p = 0.014) and was found to be more pronounced among female (χ² = 8.37, p = 0.02) than male (χ² = 6.11, p = 0.047) patients. A significant association was detected between LT-α +249A/G and increased risk of diabetes, in particular for young-onset patients (χ² = 6.92, p = 0.031). Moreover, we reported significant differences in levels of HbA1c, triglycerides, alanine transaminase, and anti-glutamic acid decarboxylase-65 among alleles. Additional studies with extended patient age groups and different ethnicities are needed to confirm our findings.     

苏姜猪OLR1基因多态性及其与肉质性状的关联分析

探讨OLR1基因在苏姜猪群内的遗传多态性,以及该基因多态对苏姜猪猪肉质性状的影响。采用PCR-RFLP技术检测OLR1基因在苏姜猪试验群体中的PstⅠ酶切遗传多态性,运用单因素方差分析方法分析了该多态位点对苏姜猪肉质性状的影响。结果发现,苏姜猪试验群体OLR1基因内含子5区域内发现一个PstⅠ酶切多态性,检测到CC、CD和DD三种基因型,多态信息含量呈现中度多态性。CC型与DD型个体的肌肉失水率、大理石纹间的差异达到显著水平(P<0.05),CD型个体用色差仪测得的b值显著高于DD型(P<0.05)。因此,检测到的OLR1基因PCR-RFLP-PstⅠ多态性与大理石纹等肉质性状存在着显著的相关关系,可以作为肉质性状候选基因在苏姜猪的持续选育中加以应用。     

形目12种鸟类线粒体细胞色素b基因序列差异及其系统发育关系

采用PCR直接测序方法首次对形目 (Charadriiformes) 12种鸟类 :蒙古沙 (Charadriusmongolus)、环颈(Charadriusalexandrinus)、大杓鹬 (Numeniusmadagascariensis)、白腰杓鹬 (Numeniusarquata)、中杓鹬 (Numeniusphaeo pus)、红脚鹬 (Tringatotanus)、林鹬 (Tringaglareola)、翘嘴鹬 (Xenuscineres)、翻石鹬 (Arenariainterpres)、大滨鹬 (Calidristenuirostris)、反嘴鹬 (Recurvirostraavosetts)和砺鹬 (Haematopusostralensis)线粒体cytb基因全序列进行测定 ,并以白鹳(Ciconiaciconia)的同源序列作为外群构建系统发生树。经比对 ,形目 12种鸟类线粒体cytb基因全序列均包括1143bp ,序列间未见有插入和缺失 ,共有 381个变异位点 ,种间序列差异值为 5 16 %～ 16 0 1%。重建的系统树将形目 12种鸟类分为 2个支系 :第 1支系包括红脚鹬、林鹬、翻石鹬、大滨鹬、翘嘴鹬、中杓鹬、大杓鹬和白腰杓鹬 ,其中红脚鹬、林鹬、翻石鹬、大滨鹬、翘嘴鹬聚为一支 ,中杓鹬、大杓鹬和白腰杓鹬聚为另一支 ;第 2支系包括蒙古沙、环颈、反嘴鹬和砺鹬 ,其中反嘴鹬与砺鹬互为姐妹群 ,然后再与属的两个种蒙古沙和环颈组成的姐妹群构成并系群。分子证据提示 :第 1支系中各属间及种间的系统关系与形态学研     

(行鸟)形目12种鸟类线粒体细胞色素b基因序列差异及其系统发育关系

采用PCR直接测序方法首次对Hang形目(Charadriiformes)12种鸟类：蒙古沙Hang(Charadrius mongolus)、环颈Hang(Charadrius alexandrinus)、大杓鹬(Numenius madagascariensis)、白腰杓鹬(Numenius arquata)、中杓鹬（Numenius phaeo-pus）、红脚鹬（Trina totanus）、林鹬（Trina alareola）、翘嘴鹬（Xenus cineres）、翻石鹬（Arenaria interpres）、大滨鹬（Calidris teruarostris)、反嘴鹬（Recurvirostra avosetts)和砺鹬（Haematopus ostralensis)线粒体cyt b基因全序列进行测定,并以白鹳（Ciconia ciconia)的同序列作为外群杓建系统发生树。经比对,Hang形目12种鸟类线粒体cyt b基因全序列均包括1143bp,序列间未见有插入和缺失,共有381个变异位点,种间序列差异值为5．16％—16．01％。重建的系统树将Hang形目12种鸟类分为2个支系：第1支系包括红脚鹬、林鹬、翻石鹬、大滨鹬、翘嘴鹬、中杓鹬、大杓鹬和白腰杓鹬,其中红脚鹬、林鹬、翻石鹬、大滨鹬、翘嘴鹬聚为一支,中杓鹬、大杓鹬和白腰杓鹬聚为另一支;第2支系包括蒙古沙Hang、环颈Hang、反嘴鹬和硕鹬,其中反嘴鹬与砺鹬互为姐妹群,然后再与Hang属的两个种蒙古沙Jamg和环Hang组成的姐妹群构成并系群。分子证据提示：第1支系中各属问及种间的系统关系与形态学研究结果相吻合;第2支系中的反嘴鹬与Hang鹬之间的亲缘关系较近,两者聚为姐妹群,提示将这两个类群合并为一个亚科——反嘴葫亚科更为合理,与码亚科共同组成Hang科,与核型研究结果和Sibley在新分类体系中将码科分为反嘴鹬亚科和Hang亚科、反嘴鹬族和砺鹬族属于反嘴鹬亚科的观点相一致。     

剑尾鱼线粒体细胞色素b基因的序列分析

目的　克隆和测定剑尾鱼 (Xiphophorushelleri)线粒体细胞色素b基因 (cytb)的全序列。方法　提取剑尾鱼肝脏的总DNA。设计合成特异引物进行PCR扩增。扩增产物经琼脂糖电泳检测、纯化后克隆到pGEM Teasyvectorsystem中的T载体上 ,筛选转化子 ,提取质粒 ,酶切鉴定。挑取重组质粒pGEM T xhcytb 11进行序列测定。结果　获得了剑尾鱼线粒体cytb基因的全序列 ,共 114 0bp。结论　用BLAST与GenBank中的线粒体DNA序列进行比较 ,显示剑尾鱼与其他鱼类的cytb基因具有较高的同源性 ;根据剑尾鱼与其他 13种鱼的cytb基因序列同源性所建立的进化树 ,与传统的分类地位基本吻合     

从细胞色素b基因序列差异分析神农架白熊的系统进化关系

神农架白熊的分类地位从它被发现后就一直存在着争议。文章测定了神农架白熊一个个体的Cyt b基因从423~1 140共718 bp的序列, 结合亚洲黑熊(Selenarctos thibetanus)、美洲黑熊(Euarctos americanus)、马来熊(Helarctos malayanus)、棕熊(Ursus arctos)、北极熊(Thalarctos maritimus)、懒熊(Melursus ursinus)、眼镜熊(Tremarctos ornatus)、大熊猫(Ailuropoda melanoleuca)、小熊猫(Ailurus fulgens)、浣熊(Procyon lotor)同源DNA序列进行比较, 分析了碱基组成频率、替代数、变异位点、简约信息位点数、转换/颠换比率、相似指数和分化指数。用邻接法( NJ)和最大节约法(MP)构建了分子系统树, 得到了基本相同的拓扑结构。结果显示, 神农架白熊与亚洲黑熊的亲缘关系最近, 与大熊猫的亲缘关系较远。     

鸡A-FABP基因多态性分析及其与脂肪性状的

以北京油鸡为试验材料,对A-FABP基因进行单核苷酸多态性(SNPs)检测和基因型与性状的关联分析。方差分析结果表明,不同基因型间腹脂率、皮脂厚、肌内脂肪含量差异极显著(P<0.01),体重在不同基因型间差异不显著(P >0.05)。由此推测,A-FABP可能为影响鸡脂肪代谢的主效基因或与主效基因相连锁。     

Genetic Diversity and Population Structure of Theileria annulata in Oman

Background

Theileriosis, caused by a number of species within the genus Theileria, is a common disease of livestock in Oman. It is a major constraint to the development of the livestock industry due to a high rate of morbidity and mortality in both cattle and sheep. Since little is currently known about the genetic diversity of the parasites causing theileriosis in Oman, the present study was designed to address this issue with specific regard to T. annulata in cattle.

Methods

Blood samples were collected from cattle from four geographically distinct regions in Oman for genetic analysis of the Theileria annulata population. Ten genetic markers (micro- and mini-satellites) representing all four chromosomes of T. annulata were applied to these samples using a combination of PCR amplification and fragment analysis. The resultant genetic data was analysed to provide a first insight into the structure of the T. annulata population in Oman.

Results

We applied ten micro- and mini-satellite markers to a total of 310 samples obtained from different regions (174 [56%] from Dhofar, 68 [22%] from Dhira, 44 [14.5%] from Batinah and 24 [8%] from Sharqia). A high degree of allelic diversity was observed among the four parasite populations. Expected heterozygosity for each site ranged from 0.816 to 0.854. A high multiplicity of infection was observed in individual hosts, with an average of 3.3 to 3.4 alleles per locus, in samples derived from Batinah, Dhofar and Sharqia regions. In samples from Dhira region, an average of 2.9 alleles per locus was observed. Mild but statistically significant linkage disequilibrium between pairs of markers was observed in populations from three of the four regions. In contrast, when the analysis was performed at farm level, no significant linkage disequilibrium was observed. Finally, no significant genetic differentiation was seen between the four populations, with most pair-wise F_ST values being less than 0.03. Slightly higher F_ST values (G_ST’ = 0.075, θ = 0.07) were detected when the data for T. annulata parasites in Oman was compared with that previously generated for Turkey and Tunisia.

Conclusion

Genetic analyses of T. annulata samples representing four geographical regions in Oman revealed a high level of genetic diversity in the parasite population. There was little evidence of genetic differentiation between parasites from different regions, and a high level of genetic diversity was maintained within each sub-population. These findings are consistent with a high parasite transmission rate and frequent movement of animals between different regions in Oman.     

Identification of a Novel Polymorphism in Bovine lncRNA ADNCR Gene and Its Association with Growth Traits

Adipocyte differentiation-associated long noncoding RNA (ADNCR) is a newly discovered lncRNA. It plays function by targeting miR-204 to significantly regulates the expression of the target SIRT1 gene in preadipocytes both at the level of mRNA and protein, thereby inhibiting adipogenesis. The tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) strategy is fast and accuracy at a negligible cost for SNP genotyping in large samples. In the study, a novel SNP g.1263T>A in intron 1 of bovine ADNCR gene was found. Herein, the T-ARMS-PCR assay was applied to detect the genotypes of the novel SNP of bovine ADNCR gene in 1017 individuals from seven cattle breeds and validated the accuracy by DNA sequencing assay of ninety animals representing three different genotypes. The concordance between two different methods was 100%. The association analysis indicated that this locus was significantly associated with the body weight (P?=?0.010), chest girth (P?=?0.014) and rump length (P?=?0.038) in Jinnan cattle, hucklebone width (P?=?0.032) in Qinchuan cattle, the cannon circumference (P?=?0.019) in Jinjiang cattle, respectively. These novel findings may be used for marker-assisted selection (MAS) and contribute to the performance of beef cattle in the future.     

CAPS and DHPLC Analysis of a Single Nucleotide Polymorphism in the Cytochrome b Gene Conferring Resistance to Strobilurins in Field Isolates of Blumeria graminis f. sp. hordei

A single nucleotide polymorphism in the wheat powdery mildew (Blumeria graminis f. sp. tritici) cytochrome b gene is responsible for resistance to inhibitors of the quinol outer binding site of the cytochrome bc₁ complex (QoI) fungicides. Analysis of a partial sequence of the cytochrome b gene from field isolates resistant and sensitive to QoI fungicides revealed the same point mutation in barley powdery mildew (B. graminis f. sp. hordei). Analysis of 118 and 40 barley powdery mildew isolates using a cleaved amplified polymorphic sequence assay and denaturing high performance liquid chromatography, respectively, confirmed that this single nucleotide polymorphism also confers resistance to QoI fungicides in barley powdery mildew.     

Polymorphism in a Microsatellite of the Acrp30 Gene and Its Association with Growth Traits in Goats

Acrp30 plays a critical role in the regulation of glucose and lipid homeostasis. In this study, polymorphism of the Acrp30 gene was detected by PCR-SSCP and DNA sequencing methods in 321 individuals from three goat breeds, and the association of Acrp30 gene polymorphism with growth traits in the three goat breeds was analyzed. A novel insert/deletion (GT)(5) microsatellite sequence was detected in the 5' flanking region of the gene. Three genotypes (AA, AB, and BB) were found in three breeds. There was moderate genetic diversity in the locus in the analyzed populations. Significant associations were observed between the genotypes of the locus and growth traits in the Boer goat population. The chest circumference of individuals with genotype BB was significantly greater than that of individuals with genotype AA.     

高山美利奴羊INHA基因多态性生物信息学分析及与产羔数关联分析

本研究旨在研究高山美利奴羊INHA基因外显子多态性及其与产羔数的相关性。本实验通过比对高山美利奴羊全基因组测序结果中不同个体INHA外显子,分析高山美利奴羊INHA基因的单核苷酸多态性,应用生物信息学软件分析高山美利奴羊INHA基因突变前后不同等位基因的m RNA二级结构、蛋白质的二级结构及三级结构。通过上述分析,将引起高山美利奴羊INHA编码氨基酸变化的位点作为该基因的特异性候选位点,通过直接测序法分析高山美利奴羊特异性候选位点INHA基因多态性,并分析其与产羔数的相关性。结果发现,高山美利奴羊INHA基因外显子区域存在3个SNPs,分别为T206A (Met→Lys)、T387A(Thr→Thr)、G900A (Pro→Pro);INHA基因3个SNPS都改变了RNA的最小自由能以及二级结构,错义突变T206A (Met→Lys)引起蛋白质二级结构和三级结构的改变;高山美利奴羊INHA基因T206A突变表现出3种基因型分别命名为TT、TA、AA,基因型与产羔数关联分析发现TT、TA基因型个体的产羔数极显著高于TT基因型个体(p<0.01)。本研究初步表明INHA基因是影响高山美利奴羊产羔数的一个主效基因。