北京鸭线粒体基因组全序列测定和分析

线粒体DNA作为遗传标记,已在家鸡(Gallus gallus)和家鹅(Anser anser)的研究中取得了重大进展,而对家鸭(Anas platyrhychos domesticus)的研究却很少.本研究参照近源物种线粒体基因组序列设计15对引物,通过PCR扩增、测序、拼接,获得北京鸭(A.platyrhychos)线粒体基因组全序列,初步分析其特点和各基因的定位.结果显示,北京鸭线粒体基因组全长16 604 bp,碱基组成为29.19%A、22.20%T、15.80%G、32.81%C,包含13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个非编码控制区(D-loop),基因组成及排列顺序与其他鸟类相似.基于线粒体D-loop区全序列,用N-J法构建了7种雁形目鸟类系统进化树,结果表明,北京鸭与绿头鸭(A.platyrhychos)系统进化关系较近.     

藏鸡和两种低地鸡线粒体全基因组序列 的测定和比对

藏鸡生活在青藏高原, 它对高原低氧具有一定的遗传适应能力. 丝羽乌骨鸡和寿光鸡都是低地鸡种, 缺乏对低氧环境的遗传适应能力. 鸡的线粒体基因组总长16785 bp, 共编码37个基因, 这些基因的编码产物均与线粒体的呼吸作用和氧化磷酸化相关, 它们的突变均有可能影响线粒体的功能, 因此均可被选为藏鸡低氧遗传适应的候选基因. 测定比较藏鸡和低地鸡的线粒体基因组, 可以为进一步研究藏鸡的低氧遗传适应机理提供线索. 本研究测定和比较了藏鸡、丝羽乌骨鸡和寿光鸡的线粒体全基因组序列, 发现藏鸡线粒体基因组总长为16784~16786 bp, 丝羽乌骨鸡为16785 bp, 寿光鸡为16784 bp; 比对后共发现突变(包括单核苷酸多态性(SNP)、单碱基缺失和插入)120个, 其中tRNA基因突变4个, rRNA基因突变10个, D-LOOP区突变39个, 基因间隔区突变1个, 编码蛋白质亚基的碱基突变66个(包括同义突变48个, 错义突变18个).     

藏鸡和两种低地鸡线粒体全基因组序列 的测定和比对

藏鸡生活在青藏高原, 它对高原低氧具有一定的遗传适应能力. 丝羽乌骨鸡和寿光鸡都是低地鸡种, 缺乏对低氧环境的遗传适应能力. 鸡的线粒体基因组总长16785 bp, 共编码37个基因, 这些基因的编码产物均与线粒体的呼吸作用和氧化磷酸化相关, 它们的突变均有可能影响线粒体的功能, 因此均可被选为藏鸡低氧遗传适应的候选基因. 测定比较藏鸡和低地鸡的线粒体基因组, 可以为进一步研究藏鸡的低氧遗传适应机理提供线索. 本研究测定和比较了藏鸡、丝羽乌骨鸡和寿光鸡的线粒体全基因组序列, 发现藏鸡线粒体基因组总长为16784~16786 bp, 丝羽乌骨鸡为16785 bp, 寿光鸡为16784 bp; 比对后共发现突变(包括单核苷酸多态性(SNP)、单碱基缺失和插入)120个, 其中tRNA基因突变4个, rRNA基因突变10个, D-LOOP区突变39个, 基因间隔区突变1个, 编码蛋白质亚基的碱基突变66个(包括同义突变48个, 错义突变18个).     

赤麂线粒体全基因组的序列和结构

提取赤麂细胞株总DNA,参照我们实验室已测定的同属动物小麂线粒体全基因组序列设计引物,PCR扩增、测序、拼接,获得赤麂线粒体全基因组序列并进行生物信息学分析。赤麂线粒体全基因组序列全长16354bp。定位了22个tRNA基因、2个rRNA基因、13个蛋白编码基因和1个D-loop区。赤麂与小麂及其它哺乳动物线粒体的基因组结构相同,它们的序列同源性都较高。     

日本条螽线粒体基因组序列的测定与分析

日本条螽完整的线粒体基因组序列长16 281 bp,包括13个蛋白质编码基因、22个tRNA基因、2个r RNA基因和1个D-loop区,其基因次序和方向与祖先序列相同。该线粒体基因组排列紧凑,但在ND2和tRNA~(Trp)之间有一段长为650 bp的基因间隔区。为研究螽斯科的系统发育关系,本研究选取日本条螽及其它17个螽斯科物种线粒体基因组的蛋白质编码基因和r RNA基因序列构建贝叶斯系统发生树。     

孟加拉笛鲷线粒体基因组序列结构及其进化

采用Long-PCR扩增线粒体全基因组方法得到了孟加拉笛鲷线粒体基因组全序列.序列分析结果表明,孟加拉笛鲷线粒体基因组序列全长16 511 bp,共有13个编码蛋白质基因、22个tRNA基因、2个rRNA基因和1个D-loop区.在编码蛋白质基因中,除COⅠ是以GTG作为起始密码子外,其它均是以ATG起始,NDⅠ、COⅡ、ND3以TAG作为终止密码子,而ND4、Cyt b则以不完全的T为终止密码子,其余8个蛋白质基因的终止密码子均为TAA.孟加拉笛鲷线粒体基因组各基因长度、位置与典型的脊椎动物相似,其编码蛋白质基因和rRNA基因与其它硬骨鱼类具有很高的同源性.基于14种笛鲷线粒体区段COⅠ、COⅡ和Cyt b基因的全序列合并成的一个组合数据集构建系统进化树,显示孟加拉笛鲷与四带笛鲷关系最为密切.     

中华鳖线粒体基因组序列分析

参照近源物种线粒体基因组序列,设计17对特异引物,采用PCR产物直接测序法测得中华鳖线粒体基因组全序列.初步分析其基因组特点和各基因的定位,用pDRAW32软件预测12种限制性酶对其的酶切图谱.结果表明,中华鳖线粒体基因组全长17364bp,核苷酸组成为35.23%A、27.26%T、25.73%C、11.78%G,包括13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个非编码控制区.基于线粒体基因组编码的13个蛋白质的氨基酸序列,用NJ法和MP法构建系统进化树,分析6种龟鳖类动物之间的亲缘关系,与传统的系统分类基本一致,初步确定淡水龟科与海龟科的亲缘关系比与龟科的亲缘关系要近.     

新西兰白兔线粒体基因组全序列的测定与分析

本研究旨在获得新西兰白兔(New Zealand white rabbit)线粒体DNA基因组全序列(mtDNA).根据GenBank已经公布的近缘物种穴兔mtDNA全基因组序列(GenBank登录号:AJ001588.1),设计12对可覆盖新西兰白兔mtDNA全序列的引物,通过PCR扩增、测序、拼接,获得新西兰白兔线粒体全序列,并分析其特点.新西兰白兔线粒体基因组全序列为17 418 bp,A+T含量高,为59.72％,蛋白编码基因数量为13个,rRNA基因数量为2个,tRNA基因数量为22个和1个非编码控制区(D-loop区),与其他兔属动物线粒体全基因组排列顺序一致.分析4种特殊的tRNA二级结构,发现tRNA-Ser(AGY)为二叶草型,缺失DHU臂,其余三种tRNA均为三叶草型.与其他哺乳动物线粒体基因组的D-loop区相比,新西兰白兔与格拉达野兔(Le-pus granatensis)核苷酸组成、编码偏好性和氨基酸组成较为相近.相比穴兔的线粒体基因组,新西兰白兔具有一定的保守性和异质性,该结果为其遗传种质资源保护和利用提供基础资料.     

小麂线粒体基因组全序列的测定和分析

通过建立麂属动物小麂线粒体DNA文库、鸟枪法测序,获得了小麂线粒体基因组全序列并对其基因组成、蛋白质的编码序列、tRNA基因等结构作了详细分析,这也是国内有关哺乳动物线粒体基因组全序列的首次报道。与其他哺乳动物线粒体基因组全序列的比较研究发现：全长为16 354bp的小麂线粒体基因组同样编码13种蛋白质、2种rRNA和22种tRNA,除了用于调控线粒体DNA复制和转录的D-Loop区以外,小麂线粒体基因组各基因长度、位置与其他哺乳动物相似,其编码蛋白质区域和rRNA基因与其他哺乳动物具有很高的同源性。     

鳙的线粒体基因组核苷酸全序列分析

对采集自我国长江的鳙的线粒体DNA全序列进行了测定.结果表明,鳙的线粒体DNA全长为166221 bp,其碱基因组成为A=31.6%;C=27.1%;G=16.0%;T=25.3%,A+T含量为56.9%.鳙线粒体基因组的排列、结构和组成与其它鲤科鱼类相似,包括37个基因,即13个蛋白质编码基因,2个rRNA基因,22个tRNA基因和一个非编码控制区(D-loop).在13个蛋白编码基因中,除ND6由轻链编码外,其余12个基因均由重链编码.COI基因的起始密码子为GTG,而其它12个蛋白编码基因的起始密码子均为ATG.     

Sequencing and alignment of mitochondrial genomes of Tibetan chicken and two lowland chicken breeds

Tibetan chicken lives in high-altitude area and has adapted well to hypoxia genetically. Shouguang chicken and Silky chicken are both lowland chicken breeds. In the present study, the complete mitochondrial genome sequences of the three chicken breeds were all sequenced. The results showed that the mitochondrial DNAs (mtDNAs) of Shouguang chicken and Silky chicken consist of 16784 bp and 16785 bp respectively, and Tibetan chicken mitochondrial genome varies from 16784 bp to 16786 bp. After sequence analysis, 120 mutations, including 4 single nucleotide polymorphisms (SNPs) in tRNA genes, 9 SNPs and 1 insertion in rRNA genes, 38 SNPs and 1 deletion in D-LOOP, 66 SNPs in protein-coding genes, were found. This work will provide clues for the future study on the association between mitochondrial genes and the adaptation to hypoxia.     

Sequencing and alignment of mitochondrial genomes of Tibetan chicken and two lowland chicken breeds

Tibetan chicken lives in high-altitude area and has adapted well to hypoxia genetically. Shouguang chicken and Silky chicken are both lowland chicken breeds. In the present study, the complete mito-chondrial genome sequences of the three chicken breeds were all sequenced. The results showed that the mitochondrial DNAs (mtDNAs) of Shouguang chicken and Silky chicken consist of 16784 bp and 16785 bp respectively, and Tibetan chicken mitochondrial genome varies from 16784 bp to 16786 bp. After sequence analysis, 120 mutations, including 4 single nucleotide polymorphisms (SNPs) in tRNA genes, 9 SNPs and 1 insertion in rRNA genes, 38 SNPs and 1 deletion in D-LOOP, 66 SNPs in pro-tein-coding genes, were found. This work will provide clues for the future study on the association between mitochondrial genes and the adaptation to hypoxia.Tibetan chicken, lowland chicken, mitochondrial genome, hypoxia.     

The mitochondrial genome of <Emphasis Type="Italic">Protohermes concolorus</Emphasis> Yang et Yang 1988 (Insecta: Megaloptera: Corydalidae)

The first complete mitochondrial genome of dobsonfly Protohermes concolorus Yang et Yang, 1988 (Megaloptera: Corydalidae) was sequenced in this study. The genome was a circular molecule of 15,851 bp containing the typical 37 genes that arranged in the same order as that of the putative ancestor of hexapods. Sequences overlaps were observed between several neighbor genes, which made the genome relatively compact. The tRNA-Ser (GCT) could not be folded into typical secondary structure because its DHU arm was replaced with a simple loop. Six of the 13 protein genes were terminated with a single T adjacent to a downstream tRNA gene in the same strand. The variation of GC content caused the different nucleotide substitution patterns of the protein genes. The genome was AT-biased with a total A + T content of 75.83% which was also demonstrated by the codon usage. The control region was the most AT-rich region with a sub-region of even higher A + T content. Protein genes of two strands presented opposite CG-skew trends which was also reflected by the codon usage. For most of the amino acids, the protein coding sequences did not prefer to use the cognate codons of corresponding tRNAs and the codon usage of the protein genes was not random. The variation of nucleotide substitution patterns of protein genes was significantly correlated with the GC content. The phylogenetic analyses based on all the 13 protein genes showed that Megaloptera was the sister group of other holometabolous insects except Coleoptera.     

Complete mitochondrial genome sequence from an endangered Indian snake, Python molurus molurus (Serpentes, Pythonidae)

This paper reports the complete mitochondrial genome sequence of an endangered Indian snake, Python molurus molurus (Indian Rock Python). A typical snake mitochondrial (mt) genome of 17258 bp length comprising of 37 genes including the 13 protein coding genes, 22 tRNA genes, and 2 ribosomal RNA genes along with duplicate control regions is described herein. The P. molurus molurus mt. genome is relatively similar to other snake mt. genomes with respect to gene arrangement, composition, tRNA structures and skews of AT/GC bases. The nucleotide composition of the genome shows that there are more A-C % than T-G% on the positive strand as revealed by positive AT and CG skews. Comparison of individual protein coding genes, with other snake genomes suggests that ATP8 and NADH3 genes have high divergence rates. Codon usage analysis reveals a preference of NNC codons over NNG codons in the mt. genome of P. molurus. Also, the synonymous and non-synonymous substitution rates (ka/ks) suggest that most of the protein coding genes are under purifying selection pressure. The phylogenetic analyses involving the concatenated 13 protein coding genes of P. molurus molurus conformed to the previously established snake phylogeny.     

甜果螨全线粒体基因组测序与分析

【目的】测定和分析甜果螨Carpoglyphus lactis线粒体基因组全序列,并在线粒体基因组水平探讨其在真螨总目(Acariformes)中的系统发育地位,为真螨总目分类及果螨科线粒体基因组研究提供科学依据。【方法】挑取实验室饲养的甜果螨成螨,用传统的酚氯仿抽提法和试剂盒提取法提取甜果螨基因组DNA。然后采用节肢动物或螨类线粒体基因的通用引物PCR扩增出甜果螨线粒体基因cox1,cob,rrnS和nad4-nad5的部分序列;再设计种特异性引物进行Long-PCR扩增和步移法测序,测出甜果螨线粒体基因组全序列。应用SeqMan, SEQUIN 9.0和tRNAscan等生物信息学软件,对甜果螨线粒体基因组的基因结构等进行生物信息学分析。最后基于17种真螨总目螨类的蛋白质编码基因,采用最大似然法构建系统发育树。【结果】甜果螨线粒体全基因组总长为14 060 bp(GenBank登录号:MN073839),为典型的闭合双链DNA分子,共由37个基因组成,包括13个蛋白质编码基因(PCGs)、22个tRNA基因和2个rRNA基因;甜果螨线粒体基因组还包括1个大的非编码区(large non-coding region, LNR)。系统发育分析结果显示,甜果螨Carpoglyphus lactis属于无气门亚目粉螨总科(Acaroidae),与椭圆食粉螨Aleuroglyphus ovatus构成一支。粉螨总科(Acaroidae)和薄口螨总科(Histiostomatoidae)聚成一簇,与痒螨股(Psoroptidia)构成姐妹群。【结论】本研究首次获得并分析了甜果螨线粒体基因组全序列。甜果螨与椭圆食粉螨的亲缘关系较近。     

Sequence and gene organization of the chicken mitochondrial genome. A novel gene order in higher vertebrates

The 16,775 base-pair mitochondrial genome of the white Leghorn chicken has been cloned and sequenced. The avian genome encodes the same set of genes (13 proteins, 2 rRNAs and 22 tRNAs) as do other vertebrate mitochondrial DNAs and is organized in a very similar economical fashion. There are very few intergenic nucleotides and several instances of overlaps between protein or tRNA genes. The protein genes are highly similar to their mammalian and amphibian counterparts and are translated according to the same variant genetic code. Despite these highly conserved features, the chicken mitochondrial genome displays two distinctive characteristics. First, it exhibits a novel gene order, the contiguous tRNA(Glu) and ND6 genes are located immediately adjacent to the displacement loop region of the molecule, just ahead of the contiguous tRNA(Pro), tRNA(Thr) and cytochrome b genes, which border the displacement loop region in other vertebrate mitochondrial genomes. This unusual gene order is conserved among the galliform birds. Second, a light-strand replication origin, equivalent to the conserved sequence found between the tRNA(Cys) and tRNA(Asn) genes in all vertebrate mitochondrial genomes sequenced thus far, is absent in the chicken genome. These observations indicate that galliform mitochondrial genomes departed from their mammalian and amphibian counterparts during the course of evolution of vertebrate species. These unexpected characteristics represent useful markers for investigating phylogenetic relationships at a higher taxonomic level.     

Mitochondrial genome of Plateau zokor Myospalax baileyi

Plateau zokors Myospalax baileyi are subterranean rodents and endemic to the cold and hypoxic (2800-4300 m above sea level) climate of the farm, prairie, alpine prairie, and meadow habitats across the Tibetan plateau. The complete mitochondrial (mt) genome of M. baileyi has been determined. Our results showed that the total length of the mitogenome was 16,351 bp, and had a gene content of 13 protein coding, 22 tRNAs and 2 rRNAs. Except for the seven tRNA and Nd6 genes, all other mt genes are encoded on the heavy strand. The overall base composition of the heavy strand is 33.65% A, 29.65% T, 24.42% C, and 12.28% G, with an AT content of 63.3%.     

绿环缘蝽(昆虫纲,半翅目,缘蝽科)线粒体基因组分析(英文)

本研究测定了昆虫纲半翅几姬缘蝽科Rhopalidae绿环缘蝽Stictopleurussubviridis Hsiao的完整线粒体基凶组.为异翅亚目蝽次目Penmtomomorha中的首例研究.该线粒体基因组为全长15 139 bp的双链环状DNA分子,含有后生动物线粒体基凶组中典型的37个基因,AT含量为75.7%.各基因的排列方式与六足动物假想祖先相同.该线粒体基因组结构紧凑,有基因重叠现象.在tRNA-Ⅱe与tRNA-GLn问发现一个罕见的79bp的非编码序列.tRNA-Ser的二级结构中DHU臂缺失,无法折叠成典型的三叶草结构.CO2和CO3终止密码子为T,其下游为同链编码的tRNA基因.基因组AT含量为75.7%,密码子使用频率也反映出AT偏好.两条链上蛋白质编码基凶的CG-skew趋势相反,密码子使用频率也反映出两链不同的CG偏好.蛋白编码序列并不总是使用与tRNA的反密码子相对应的密码子,密码子的使用并不随机.使用频率最高密码子为NNA,使用频率最低的密码子为NNG.与臭虫次目Cimicomomorpha中的锥猎蝽Triaatoma dimidiata线粒体基因组比较分析表明,ATP8为进化最快的蛋白质,CO1最慢,但tRNA和rRNA基因的进化速率比蛋白质基因低.