ITS序列在天门冬属物种DNA条形码构建中的应用研究

在对天门冬属4个种ITS(internal transcribed spacer)序列克隆测序的基础上,并对4个种进行序列比较分析.结果表明:(1)4个物种的亲缘关系很近,它们间的ITS序列的变异小,GC含量高,在60％左右.(2)4个物种在系统树上聚为一类,文竹与其他3种天门冬的亲缘关系相对较远,处于系统树的一支,其...     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County ( Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta . In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

Gene regulation: ancient microRNA target sequences in plants

MicroRNAs are an abundant class of small RNAs that are thought to regulate the expression of protein-coding genes in plants and animals. Here we show that the target sequence of two microRNAs, known to regulate genes in the class-III homeodomain-leucine zipper (HD-Zip) gene family of the flowering plant Arabidopsis, is conserved in homologous sequences from all lineages of land plants, including bryophytes, lycopods, ferns and seed plants. We also find that the messenger RNAs from these genes are cleaved within the same microRNA-binding site in representatives of each land-plant group, as they are in Arabidopsis. Our results indicate not only that microRNAs mediate gene regulation in non-flowering as well as flowering plants, but also that the regulation of this class of plant genes dates back more than 400 million years.     

Conservation and change in the DNA sequences coding for alcohol dehydrogenase in sibling species of Drosophila

The DNA sequences of the alcohol dehydrogenase (Adh) genes of four very closely related species of Drosophila show that the rates of nucleotide change vary greatly in different functional domains of this gene. A phylogeny of these species based on the Adh sequence data is consistent with that based on polytene chromosome banding patterns.     

核酸分子“光开关”作探针荧光光谱法检测转基因启动子病毒DNA特定序列

合成核酸分子"光开关"Ru(phen2)(dppx)2+,并以其作为荧光探针检测转基因花椰菜花叶病毒35S启动子(CaMV 35S)序列片段(5’-CATCGTT GAAGA TGCCTCTGCC G-3’).通过对检测条件的优化,测定CaMV 35S病毒DNA的线性范围为1.4×10-9～6.9×10-8 mol.L-1,线性关系:ΔI=1.6913C+5.5783(式中ΔI为标准溶液的荧光强度与空白溶液的荧光强度之差),R2=0.9938,检出限为8.6×10-10 mol/L.实验表明该方法有操作简单、灵敏度高和选择性好等特点,同时可以较好地识别碱基错配序列和非互补序列ssDNA.     

利用定积分定义求数列极限

讨论了应用定积分定义求数列极限的方法 ,并给出了确定被积函数及积分上、下限的具体步骤     

不同物种间胰岛素及其编码mRNA,DNA序列比较与分析

通过PSI-BLAST搜索与人类胰岛素原(含有86个氨基酸)相似的蛋白质序列，并进行比对，计算比对矩阵的相似得分和期望值，同时运用ClustalW算法对不同物种编码前胰岛素原mRNA及其翻译的蛋白质和DNA序列进行多重比对．结果发现，脊椎动物的胰岛素蛋白质一级结构(A链和B链)和mRNA非常相似，但部分动物C肽的部分序列有差异；系统进化分析表明，人和猴、小鼠和大鼠编码胰岛素的mRNA在进化上关系相近．各物种间编码相同氨基酸的核苷酸序列（CDS)相同，但编码胰岛素的DNA序列不同．各物种胰岛素原蛋白质序列中，A链和B链序列保守，C肽有一定的差异；DNA序列差异较大．     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County (Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta. In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

New ancient DNA sequences suggest high genetic diversity for the woolly mammoth (Mammuthus primigenius)

Partial DNA sequences of cytochrome b gene (mtDNA) were successfully retrieved from Late Pleistocene fossil bone of Mammuthus primigenius collected from the Xiguitu County (Yakeshi), Inner Mongolia Autonomous Region and from Zhaodong, Harbin of Heilongjiang Province in northern China. Two ancient DNA fragments (109 bp and 124 bp) were authenticated by reproducible experiments in two different laboratories and by phylogenetic analysis with other Elephantidae taxa. Phylogenetic analysis using these sequences and published data in either separate or combined datasets indicate unstable relationship among the woolly mammoth and the two living elephants, Elephas and Loxodonta. In addition to the short sequences used to attempt the long independent evolution of Elephantidae terminal taxa, we suggest that a high intra-specific diversity existed in Mammuthus primigenius crossing both spatial and temporal ranges, resulting in a complex and divergent genetic background for DNA sequences so far recovered. The high genetic diversity in the extinct woolly mammoth can explain the apparent instability of Elephantidae taxa on the molecular phylogenetic trees and can reconcile the apparent paradox regarding the unresolved Elephantidae trichotomy.     

Mitochondrial DNA analysis of ancient Sampula population in Xinjiang

The archaeological site of Sampula cemetery was located about 14 km to the southwest of the Luo County in Xinjiang Khotan,China,belonging to the ancient Yutian kingdom.~14C analysis showed that this cemetery was used from 217 B.C.to 283 A.D. Ancient DNA was analyzed by 364 bp of the mitochondrial DNA hypervariable region I (mtDNA HVR-I),and by six restriction fragment length polymorphism (RFLP) sites of mtDNA coding region.We successfully extracted and sequenced intact stretches of maternally inher- ited mtDNA from 13 out of 16 ancient Sampula samples.The analysis of mtDNA haplogroup distribution showed that the ancient Sampula was a complex population with both European and Asian characteristics.Median joining network of U3 sub-haplogroup and multi-dimen- sional scaling analysis all showed that the ancient Sampula had maternal relationship with Ossetian and Iranian.     

Mitochondrial DNA analysis of ancient Sampula population in Xinjiang

The archaeological site of Sampula cemetery was located about 14 km to the southwest of the Luo County in Xinjiang Khotan, China, belonging to the ancient Yutian kingdom. ¹⁴C analysis showed that this cemetery was used from 217 B.C. to 283 A.D. Ancient DNA was analyzed by 364 bp of the mitochondrial DNA hypervariable region I (mtDNA HVR-I), and by six restriction fragment length polymorphism (RFLP) sites of mtDNA coding region. We successfully extracted and sequenced intact stretches of maternally inherited mtDNA from 13 out of 16 ancient Sampula samples. The analysis of mtDNA haplogroup distribution showed that the ancient Sampula was a complex population with both European and Asian characteristics. Median joining network of U3 sub-haplogroup and multi-dimensional scaling analysis all showed that the ancient Sampula had maternal relationship with Ossetian and Iranian.     

DNA sequences of telomeres maintained in yeast

Telomeres, the ends of eukaryotic chromosomes, have long been recognized as specialized structures. Their stability compared with broken ends of chromosomes suggested that they have properties which protect them from fusion, degradation or recombination. Furthermore, a linear DNA molecule such as that of a eukaryotic chromosome must have a structure at its ends which allows its complete replication, as no known DNA polymerase can initiate synthesis without a primer. At the ends of the relatively short, multi-copy linear DNA molecules found naturally in the nuclei of several lower eukaryotes, there are simple tandemly repeated sequences with, in the cases analysed, a specific array of single-strand breaks, on both DNA strands, in the distal portion of the block of repeats. In general, however, direct analysis of chromosomal termini presents problems because of their very low abundance in nuclei. To circumvent this problem, we have previously cloned a chromosomal telomere of the yeast Saccharomyces cerevisiae on a linear DNA vector molecule. Here we show that yeast chromosomal telomeres terminate in a DNA sequence consisting of tandem irregular repeats of the general form C1-3A. The same repeat units are added to the ends of Tetrahymena telomeres, in an apparently non-template-directed manner, during their replication on linear plasmids in yeast. Such DNA addition may have a fundamental role in telomere replication.     

Cortical growth marks reveal extended juvenile development in New Zealand moa

Cyclical growth marks in cortical bone, deposited before attainment of adult body size, reflect osteogenetic changes caused by annual rhythms and are a general phenomenon in non-avian ectothermic and endothermic tetrapods. However, the growth periods of ornithurines (the theropod group including all modern birds) are usually apomorphically shortened to less than a year, so annual growth marks are almost unknown in this group. Here we show that cortical growth marks are frequent in long bones of New Zealand's moa (Aves: Dinornithiformes), a recently extinct ratite order. Moa showed the exaggerated K-selected life-history strategy formerly common in the New Zealand avifauna, and in some instances took almost a decade to attain skeletal maturity. This indicates that reproductive maturity in moa was extremely delayed relative to all extant birds. The two presently recognized moa families (Dinornithidae and Emeidae) also showed different postnatal growth rates, which were associated with their relative differences in body size. Both species of giant Dinornis moa attained their massive stature (up to 240 kg live mass) by accelerating their juvenile growth rate compared to the smaller emeid moa species, rather than by extending the skeletal growth period.     

Phylogenetic position of Ephedra rhytidosperma, a species endemic to China： Evidence from chloroplast and ribosomal DNA sequences

The chloroplast genes matK and rbcL, ribosomal gene 18S and ITS regions of nuclear ribosomal DNA from Ephedra rhytidosperma, a species endemic to China, were sequenced and its phylogenetic position was investigated. Independent and combined phylogenetic analyses for the DNA sequences from 16 taxa representing 15 species of the genus Ephedra were performed using the maximum parsimony （MP）, neighbor-joining （N J）, minimum evolution （ME） and maximum likelihood （ML） methods. The results indicate that E. rhytidosperma is closely related to E. equisetina. The divergence time between them is estimated to be 10.85±2.44 Ma based on the results of the relative-rate tests and the evolutionary rate of rbcL gene.     

Extreme reversed sexual size dimorphism in the extinct New Zealand moa Dinornis

The ratite moa (Aves; Dinornithiformes) were massive graviportal browsers weighing up to 250 kg (ref. 1) that dominated the New Zealand biota until their extinction approximately 500 yr ago. Despite an extensive Quaternary fossil record, moa taxonomy remains problematic and currently 11 species are recognized. Three Dinornis species were found throughout New Zealand and differed markedly in size (1-2 m height at back) and mass (from approximately 34 to 242 kg). Surprisingly, ancient mitochondrial DNA sequences show that the three species were genetically indistinguishable within each island, but formed separate North and South Island clades. Here we show, using the first sex-linked nuclear sequences from an extinct species, that on each island the three morphological forms actually represent just one species, whose size varied markedly according to sex and habitat. The largest females in this example of extreme reversed sexual size dimorphism were about 280% the weight and 150% the height of the largest males, which is unprecedented among birds and terrestrial mammals. The combination of molecular and palaeontological data highlights the difficulties of analysing extinct groups, even those with detailed fossil records.     

Palindromic sequences in heteroduplex DNA inhibit mismatch repair in yeast

Although single heterozygous markers in yeast usually segregate during meiosis in a 2:2 ratio, abberant 3:1 segregations occur quite frequently as a result of gene-conversion events. A second type of aberrant segregation, post-meiotic segregation, results from the segregation of two genotypes from a single haploid spore; in yeast such events are detected as sectored spore colonies and usually occur rarely. Post-meiotic segregation is thought to result from the replication of heteroduplex DNA formed during meiotic recombination. We report here that if the heteroduplex includes a palindromic insertion sequence, a high frequency of post-meiotic segregation results. This suggests that palindromic insertions are poorly repaired, which may be the result of hairpin-loop formation that affects the efficiency of repair of heteroduplex DNA.