大熊猫DNA序列变异及其遗传多样性研究

测定了来自马边、美姑、越西、宝兴、平武、青川、南坪和白水江等地40只大熊猫336～444bp的线粒体tRNA基因和D环区序列.在21个创立者中共检出9种线粒体DNA单倍型.发现大熊猫的遗传分化程度很低,地理群体间无显著的遗传隔离.大熊猫现生群体可能源于晚更新世,并受瓶颈效应的影响.在此之后,其遗传多样性得到了一定程度的恢复.     

Physical organization of the major duplication on Brassica oleracea chromosome O6 revealed through fluorescence in situ hybridization with Arabidopsis and Brassica BAC probes.

The close relationship between Brassica oleracea and Arabidopsis thaliana has been used to explore the genetic and physical collinearity of the two species, focusing on an inverted segmental chromosome duplication within linkage group O6 of B. oleracea. Genetic evidence suggests that these segments share a common origin with a region of Arabidopsis chromosome 1. Brassica oleracea and Arabidopsis bacterial artificial chromosome probes have been used for fluorescence in situ hybridization analysis of B. oleracea pachytene chromosomes to further characterize the inverted duplication. This has been highly effective in increasing the local resolution of the cytogenetic map. We have shown that the physical order of corresponding genetic markers is highly conserved between the duplicated regions in B. oleracea and the physical lengths of the regions at pachytene are similar, while the genetic distances are considerably different. The physical marker order is also well conserved between Arabidopsis and B. oleracea, with only one short inversion identified. Furthermore, the relative physical distances between the markers in one segment of B. oleracea and Arabidopsis have stayed approximately the same. The efficacy of using fluorescence in situ hybridization, together with other forms of physical and genetic mapping, for elucidating such issues relating to synteny is discussed.     

Hemagglutinin Stalk Immunity Reduces Influenza Virus Replication and Transmission in Ferrets

We assessed whether influenza virus hemagglutinin stalk-based immunity protects ferrets against aerosol-transmitted H1N1 influenza virus infection. Immunization of ferrets by a universal influenza virus vaccine strategy based on viral vectors expressing chimeric hemagglutinin constructs induced stalk-specific antibody responses. Stalk-immunized ferrets were cohoused with H1N1-infected ferrets under conditions that permitted virus transmission. Hemagglutinin stalk-immunized ferrets had lower viral titers and delayed or no virus replication at all following natural exposure to influenza virus.     

Sequence evolution of the CCR5 chemokine receptor gene in primates.

The chemokine receptor CCR5 can serve as a coreceptor for M-tropic HIV-1 infection and both M-tropic and T-tropic SIV infection. We sequenced the entire CCR5 gene from 10 nonhuman primates: Pongo pygmaeus, Hylobates leucogenys, Trachypithecus francoisi, Trachypithecus phayrei, Pygathrix nemaeus, Rhinopithecus roxellanae, Rhinopithecus bieti, Rhinopithecus avunculus, Macaca assamensis, and Macaca arctoides. When compared with CCR5 sequences from humans and other primates, our results demonstrate that: (1) nucleotide and amino acid sequences of CCR5 among primates are highly homologous, with variations slightly concentrated on the amino and carboxyl termini; and (2) site Asp13, which is critical for CD4-independent binding of SIV gp120 to Macaca mulatta CCR5, was also present in all other nonhuman primates tested here, suggesting that those nonhuman primate CCR5s might also bind SIV gp120 without the presence of CD4. The topologies of CCR5 gene trees constructed here conflict with the putative opinion that the snub-nosed langurs compose a monophyletic group, suggesting that the CCR5 gene may not be a good genetic marker for low-level phylogenetic analysis. The evolutionary rate of CCR5 was calculated, and our results suggest a slowdown in primates after they diverged from rodents. The synonymous mutation rate of CCR5 in primates is constant, about 1.1 x 10(-9) synonymous mutations per site per year. Comparisons of Ka and Ks suggest that the CCR5 genes have undergone negative or purifying selection. Ka/Ks ratios from cercopithecines and colobines are significantly different, implying that selective pressures have played different roles in the two lineages.     

Postexercise glucose uptake and glycogen synthesis in skeletal muscle from GLUT4-deficient mice.

To determine the role of GLUT4 on postexercise glucose transport and glycogen resynthesis in skeletal muscle, GLUT4-deficient and wild-type mice were studied after a 3 h swim exercise. In wild-type mice, insulin and swimming each increased 2-deoxyglucose uptake by twofold in extensor digitorum longus muscle. In contrast, insulin did not increase 2-deoxyglucose glucose uptake in muscle from GLUT4-null mice. Swimming increased glucose transport twofold in muscle from fed GLUT4-null mice, with no effect noted in fasted GLUT4-null mice. This exercise-associated 2-deoxyglucose glucose uptake was not accompanied by increased cell surface GLUT1 content. Glucose transport in GLUT4-null muscle was increased 1.6-fold over basal levels after electrical stimulation. Contraction-induced glucose transport activity was fourfold greater in wild-type vs. GLUT4-null muscle. Glycogen content in gastrocnemius muscle was similar between wild-type and GLUT4-null mice and was reduced approximately 50% after exercise. After 5 h carbohydrate refeeding, muscle glycogen content was fully restored in wild-type, with no change in GLUT4-null mice. After 24 h carbohydrate refeeding, muscle glycogen in GLUT4-null mice was restored to fed levels. In conclusion, GLUT4 is the major transporter responsible for exercise-induced glucose transport. Also, postexercise glycogen resynthesis in muscle was greatly delayed; unlike wild-type mice, glycogen supercompensation was not found. GLUT4 it is not essential for glycogen repletion since muscle glycogen levels in previously exercised GLUT4-null mice were totally restored after 24 h carbohydrate refeeding.-Ryder, J. W., Kawano, Y., Galuska, D., Fahlman, R., Wallberg-Henriksson, H., Charron, M. J., Zierath, J. R. Postexercise glucose uptake and glycogen synthesis in skeletal muscle from GLUT4-deficient mice.     

Characterization of TAP Ambr 250 disposable bioreactors,as a reliable scale‐down model for biologics process development

Demands for development of biological therapies is rapidly increasing, as is the drive to reduce time to patient. In order to speed up development, the disposable Automated Microscale Bioreactor (Ambr 250) system is increasingly gaining interest due to its advantages, including highly automated control, high throughput capacity, and short turnaround time. Traditional early stage upstream process development conducted in 2 ‐ 5 L bench‐top bioreactors requires high foot‐print, and running cost. The establishment of the Ambr 250 as a scale‐down model leads to many benefits in process development. In this study, a comprehensive characterization of mass transfer coefficient (k_La) in the Ambr 250 was conducted to define optimal operational conditions. Scale‐down approaches, including dimensionless volumetric flow rate (vvm), power per unit volume (P/V) and k_La have been evaluated using different cell lines. This study demonstrates that the Ambr 250 generated comparable profiles of cell growth and protein production, as seen at 5‐L and 1000‐L bioreactor scales, when using k_La as a scale‐down parameter. In addition to mimicking processes at large scales, the suitability of the Ambr 250 as a tool for clone selection, which is traditionally conducted in bench‐top bioreactors, was investigated. Data show that cell growth, productivity, metabolite profiles, and product qualities of material generated using the Ambr 250 were comparable to those from 5‐L bioreactors. Therefore, Ambr 250 can be used for clone selection and process development as a replacement for traditional bench‐top bioreactors minimizing resource utilization during the early stages of development in the biopharmaceutical industry. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:478–489, 2017     

Impact of Flooding on the Species Richness, Density and Composition of Amazonian Litter-Nesting Ants

Litter-nesting ants are diverse and abundant in tropical forests, but the factors structuring their communities are poorly known. Here we present results of the first study to examine the impact of natural variation in flooding on a highly diverse (21 genera, 77 species) litter-nesting ant community in a primary Amazonian forest. Fifty-six 3 × 3 m plots experiencing strong variation in flooding and twenty-eight 3 × 3 m terra firme plots were exhaustively searched for litter-nesting ants to determine patterns of density, species richness and species composition. In each plot, flooding, litter depth, twig availability, canopy cover, plant density, percent soil nitrogen, carbon, and phosphorus were measured. Degree of flooding, measured as flood frequency and flood interval, had the strongest impact on ant density in flooded forest. Flooding caused a linear decrease in ant abundance, potentially due to a reduction of suitable nesting sites. However, its influence on species richness varied: low-disturbance habitat had species richness equal to terra firme forest after adjusting for differences in density. The composition of ant genera and species varied among flood categories; some groups known to contain specialist predators were particularly intolerant to flooding. Hypoponera STD10 appeared to be well-adapted to highly flooded habitat. Although flooding did not appear to increase species richness or abundance at the habitat scale, low-flooding habitat contained a mixture of species found in the significantly distinct ant communities of terra firme and highly flooded habitat.
     

Horse domestication and conservation genetics of Przewalski's horse inferred from sex chromosomal and autosomal sequences

Despite their ability to interbreed and produce fertile offspring,there is continued disagreement about the genetic relationshipof the domestic horse (Equus caballus) to its endangered wildrelative, Przewalski's horse (Equus przewalskii). Analyses havediffered as to whether or not Przewalski's horse is placed phylogeneticallyas a separate sister group to domestic horses. Because Przewalski'shorse and domestic horse are so closely related, genetic datacan also be used to infer domestication-specific differencesbetween the two. To investigate the genetic relationship ofPrzewalski's horse to the domestic horse and to address whetherevolution of the domestic horse is driven by males or females,five homologous introns (a total of 3 kb) were sequenced onthe X and Y chromosomes in two Przewalski's horses and threebreeds of domestic horses: Arabian horse, Mongolian domestichorse, and Dartmoor pony. Five autosomal introns (a total of6 kb) were sequenced for these horses as well. The sequencesof sex chromosomal and autosomal introns were used to determinenucleotide diversity and the forces driving evolution in thesespecies. As a result, X chromosomal and autosomal data do notplace Przewalski's horses in a separate clade within phylogenetictrees for horses, suggesting a close relationship between domesticand Przewalski's horses. It was also found that there was alack of nucleotide diversity on the Y chromosome and highernucleotide diversity than expected on the X chromosome in domestichorses as compared with the Y chromosome and autosomes. Thissupports the hypothesis that very few male horses along withnumerous female horses founded the various domestic horse breeds.Patterns of nucleotide diversity among different types of chromosomeswere distinct for Przewalski's in contrast to domestic horses,supporting unique evolutionary histories of the two species.