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1.
本文从观察温度的影响出发,探讨了鼠肝线粒体内膜体,在琥珀酸氧化建立跨膜质子电化学梯度(ΔμH^+)时,膜脂双分子层中DPH荧光偏振值(r)的变化与膜能量偶联活性之间的相互关系。结果表明,15 ̄35℃温度内,能化引起r值变化趋势相似,r值变化速率随温度升高而增加,但与温度对r值影响相比只是在较小的范围内变动。另一方面,15 ̄30℃温度内,随温度升高质子回漏速率加快,RCR值和ADP/O比值下降,但跨 相似文献
2.
Roncero MI Di Pietro A Ruiz-Roldán MC Huertas-González MD Garcia-Maceira FI Méglecz E Jiménez A Caracuel Z Sancho-Zapatero R Hera C Gómez-Gómez E Ruiz-Rubio M González-Verdejo CI Páez MJ 《Revista iberoamericana de micología》2000,17(1):S47-S53
Fusarium oxysporum invades its host plants through the roots and colonizes the vascular system. It produces a great variety of cell-wall degrading enzymes (CWDE), such as cellulases, xylanases, pectinases and proteases. Our group has purified and characterized an endopolygalacturonase (PG1), two exopolygalacturonases (PG2 and PG3), an endoxylanase (XYL1) and an endo pectatelyase (PL1). We have isolated the following CWDE-encoding genes: pg1, pgx4, pg5, xyl2, xyl3, prt1 and pl1. Gene expression in different culture conditions has been determined by Northern analysis. The occurrence of these genes in different formae speciales has been analyzed by Southern analysis and PCR. All these genes are expressed during different stages of the interaction with the host plant indicating a possible role in pathogenesis. At present, targeted gene disruption is being carried out, in order to determine the role of each gene in the pathogenicity process. 相似文献
3.
Mark W Ronsyn Jasmijn Daans Gie Spaepen Shyama Chatterjee Katrien Vermeulen Patrick D'Haese Viggo FI Van Tendeloo Eric Van Marck Dirk Ysebaert Zwi N Berneman Philippe G Jorens Peter Ponsaerts 《BMC biotechnology》2007,7(1):1-17
Background
Swine is an important agricultural commodity and biomedical model. Manipulation of the pig genome provides opportunity to improve production efficiency, enhance disease resistance, and add value to swine products. Genetic engineering can also expand the utility of pigs for modeling human disease, developing clinical treatment methodologies, or donating tissues for xenotransplantation. Realizing the full potential of pig genetic engineering requires translation of the complete repertoire of genetic tools currently employed in smaller model organisms to practical use in pigs.Results
Application of transposon and recombinase technologies for manipulation of the swine genome requires characterization of their activity in pig cells. We tested four transposon systems- Sleeping Beauty, Tol2, piggyBac, and Passport in cultured porcine cells. Transposons increased the efficiency of DNA integration up to 28-fold above background and provided for precise delivery of 1 to 15 transgenes per cell. Both Cre and Flp recombinase were functional in pig cells as measured by their ability to remove a positive-negative selection cassette from 16 independent clones and over 20 independent genomic locations. We also demonstrated a Cre-dependent genetic switch capable of eliminating an intervening positive-negative selection cassette and activating GFP expression from episomal and genome-resident transposons.Conclusion
We have demonstrated for the first time that transposons and recombinases are capable of mobilizing DNA into and out of the porcine genome in a precise and efficient manner. This study provides the basis for developing transposon and recombinase based tools for genetic engineering of the swine genome. 相似文献4.
The endoglucanase coded by celA (GenBank Access No. Y12512) from Paenibacillus barcinonensis, an enzyme with good characteristics for application on paper manufacture from agricultural fibers, was expressed in Saccharomyces cerevisiae by using different domains of the cell wall protein Pir4 as translational fusion partners, to achieve either secretion or
cell wall retention of the recombinant enzyme. Given the presence of five potential N-glycosylation sites in the amino acid sequence coded by celA, the effect of glycosylation on the enzymatic activity of the recombinant enzyme was investigated by expressing the recombinant
fusion proteins in both, standard and glycosylation-deficient strains of S. cerevisiae. Correct targeting of the recombinant fusion proteins was confirmed by Western immunoblot using Pir-specific antibodies,
while enzymatic activity on carboxymethyl cellulose was demonstrated on plate assays, zymographic analysis and colorimetric
assays. Hyperglycosylation of the enzyme when expressed in the standard strain of S. cerevisiae did not affect activity, and values of 1.2 U/ml were obtained in growth medium supernatants in ordinary batch cultures after
24 h. These values compare quite favorably with those described for other recombinant endoglucanases expressed in S. cerevisiae. This is one of the few reports describing the expression of Bacillus cellulases in S. cerevisiae, since yeast expressed recombinant cellulases have been mostly of fungal origin. It is also the first report of the yeast
expression of this particular endoglucanase. 相似文献
5.
João Batista A Oliveira Mario Cavagna Claudia G Petersen Ana L Mauri Fabiana C Massaro Liliane FI Silva Ricardo LR Baruffi Jose G Franco Jr 《Reproductive biology and endocrinology : RB&E》2011,9(1):1-7
Background
The role of serum anti-Müllerian hormone (AMH) as predictor of in-vitro fertilization outcomes has been much debated. The aim of the present study is to investigate the practicability of combining serum AMH level with biological age as a simple screening method for counseling IVF candidates of advanced reproductive age with potential poor outcomes prior to treatment initiation.Methods
A total of 1,538 reference patients and 116 infertile patients aged greater than or equal to 40 years enrolled in IVF/ICSI cycles were recruited in this retrospective analysis. A reference chart of the age-related distribution of serum AMH level for Asian population was first created. IVF/ICSI patients aged greater than or equal to 40 years were then divided into three groups according to the low, middle and high tertiles the serum AMH tertiles derived from the reference population of matching age. The cycle outcomes were analyzed and compared among each individual group.Results
For reference subjects aged greater than or equal to 40 years, the serum AMH of the low, middle and high tertiles were equal or lesser than 0.48, 0.49-1.22 and equal or greater than 1.23 ng/mL respectively. IVF/ICSI patients aged greater than or equal to 40 years with AMH levels in the low tertile had the highest cycle cancellation rate (47.6%) with zero clinical pregnancy. The nadir AMH level that has achieved live birth was 0.56 ng/mL, which was equivalent to the 36.4th percentile of AMH level from the age-matched reference group. The optimum cut-off levels of AMH for the prediction of nonpregnancy and cycle cancellation were 1.05 and 0.68 ng/mL, respectively.Conclusions
Two criteria: (1) age greater than or equal to 40 years and (2) serum AMH level in the lowest tertile (equal or lesser than 33.3rd percentile) of the matching age group, may be used as markers of futility for counseling IVF/ICSI candidates. 相似文献6.
7.
João Batista A Oliveira Claudia G Petersen Fabiana C Massaro Ricardo LR Baruffi Ana L Mauri Liliane FI Silva Juliana Ricci José G FrancoJr 《Reproductive biology and endocrinology : RB&E》2010,8(1):56
Background
Although the motile sperm organelle morphology examination (MSOME) was developed only as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in evaluation of semen quality, with potential clinical repercussions. The present study aimed to evaluate individual variations in the motile sperm organelle morphology examination (MSOME) analysis after a time interval. 相似文献8.
Claudia G Petersen Laura D Vagnini Ana L Mauri Fabiana C Massaro Liliane FI Silva Mario Cavagna Ricardo LR Baruffi Joao BA Oliveira José G FrancoJr 《Reproductive biology and endocrinology : RB&E》2011,9(1):53
Background
This study evaluated whether there is a relationship between the zona pellucida birefringence (ZP-BF) intensity and the nuclear (NM) and cytoplasmic (CM) in vitro maturation of human oocytes from stimulated cycles. 相似文献9.
牛生长激素基因的人工合成,重组克隆及高效表达 总被引:2,自引:0,他引:2
本文通过设计选择大肠杆菌高频利用密码子代替天然密码子,人工全合成32个寡聚核苷酸片段;连接并克隆获得A(278bp)、B(88bp)、C(224bp)3个较大DNA片段;经重组克隆、定位突变等获得阳性克隆;双向DNA序列测定分析表明获得了两种人工全合成牛生长激素(bGH)编码基因,即编码N-Met-Ala-bGH和N-Met-Phe-bGH的两个基因,而至今尚未见有人工全合成bGH基因的报道。构建了在PLpromoter控制下含上述合成基因的表达质位pBLbGHE7A1和pBLbGHE8,并在大肠杆菌中进行了温敏诱导表达。经SDS-PAGE分析,表达产物占全菌蛋白的37%以上。Western-blot分析和纯化产物的N端氨基酸序列测定结果都表明上述两种人工全合成bGH基因在大肠杆菌中得到了高效表达,表达量为现今国际文献报道的高限水平。 相似文献
10.
MATJAŽ KUNTNER SIMONA KRALJ‐FIŠER MATJAŽ GREGORIČ 《Biological journal of the Linnean Society. Linnean Society of London》2010,99(4):849-866
Spider web research bridges ethology, ecology, functional morphology, material science, development, genetics, and evolution. Recent work proposes the aerial orb web as a one‐time key evolutionary innovation that has freed spider‐web architecture from substrate constraints. However, the orb has repeatedly been modified or lost within araneoid spiders. Modifications include not only sheet‐ and cobwebs, but also ladder webs, which secondarily utilize the substrate. A recent nephilid species level phylogeny suggests that the ancestral nephilid web architecture was an arboricolous ladder and that round aerial webs were derived. Because the web biology of the basalmost Clitaetra and the derived Nephila are well understood, the present study focuses on the webs of the two phylogenetically intervening genera, Herennia and Nephilengys, to establish ontogenetic and macroevolutionary patterns across the nephilid tree. We compared juvenile and adult webs of 95 Herennia multipuncta and 143 Nephilengys malabarensis for two measures of ontogenetic allometric web changes: web asymmetry quantified by the ladder index, and hub asymmetry quantified by the hub displacement index. We define a ‘ladder web’ as a vertically elongated orb exceeding twice the length over width (ladder index ≥ 2) and possessing (sub)parallel rather than round side frames. Webs in both genera allometrically grew from orbs to ladders, more so in Herennia. Such allometric web growth enables the spider to maintain its arboricolous web site. Unexpectedly, hub asymmetry only increased significantly in heavy‐bodied Nephilengys females, and not in Herennia, challenging the commonly invoked gravity hypothesis. The findings obtained in the present study support the intrageneric uniformness of nephilid webs, with Herennia etruscilla webs being identical to H. multipuncta. The nephilid web evolution suggests that the ancestor of Nephila reinvented the aerial orb web because the orb arises at a much more inclusive phylogenetic level, and all intervening nephilids retained the secondarily acquired substrate‐dependent ladder web. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 849–866. 相似文献