Crystallization of the first three domains of the human insulin-like growth factor-1 receptor.

The insulin-like growth factor-1 receptor (IGF-1R) is a tyrosine kinase receptor of central importance in cell proliferation. A fragment (residues 1-462) comprising the L1-cysteine rich-L2 domains of the human IGF-1R ectodomain has been overexpressed in glycosylation-deficient Lec8 cells and has been affinity-purified via a c-myc tag followed by gel filtration. The fragment was recognized by two anti-IGF-1R monoclonal antibodies, 24-31 and 24-60, but showed no detectable binding of IGF-1 or IGF-2. Isocratic elution of IGF-1R/462 on anion-exchange chromatography reduced sample heterogeneity, permitting the production of crystals that diffracted to 2.6 A resolution with cell dimensions a = 77.0 A, b = 99.5 A, c = 120.1 A, and space group P2(1)2(1)2(1).     

Design, synthesis, expression, and characterization of the genes for mouse Fc gamma RIIb1 and Fc gamma RIIb2 cytoplasmic regions.

The cytoplasmic regions of the mouse low-affinity Fc gamma RII isoforms, mFc gamma RIIb1, and mFc gamma RIIb2, play a key role in signal transduction by mediating different cellular functions. mFc gamma RIIb1 has a 94-residue cytoplasmic region, whereas mFc gamma RIIb2 has a 47-residue cytoplasmic region. Genes encoding the cytoplasmic regions of mFc gamma RIIb1 (b1-94) and mFc gamma RIIb2 (b2-47) were designed, synthesized, and expressed as fusion proteins in Escherichia coli. A sequence-specific protease, thrombin, was used to release the b1-94 peptide, which was purified by using HPLC. The b2-47 peptide was synthesized chemically. CD spectropolarimetry was employed to examine the secondary structures of b1-94 and b2-47. These studies were conducted in aqueous solution, in mixtures of water and trifluoroethanol or methanol, and as a function of temperature. The results indicate that the b1-94 and b2-47 structures are sensitive functions of the solvent environment, and that nonaqueous solvents induce significant alpha-helical structure.     

Population structure and impact of supportive breeding inferred from mitochondrial and microsatellite DNA analyses in land-locked Atlantic salmon Salmo salar L.

Four tributaries of Lake St-Jean (Québec, Canada) are used for spawning and juvenile habitat by land-locked Atlantic salmon. Spawning runs have drastically declined since the mid-1980s, and consequently, a supportive-breeding programme was undertaken in 1990. In this study, we analysed seven microsatellite loci and mtDNA, and empirically estimated effective population sizes to test the hypotheses that (i) fish spawning in different tributaries form genetically distinct populations and (ii) the supportive breeding programme causes genetic perturbations on wild populations. Allele frequency distribution, molecular variance and genetic distance estimates all supported the hypothesis of genetic differentiation among salmon from different tributaries. Gene flow among some populations was much more restricted than previously reported for anadromous populations despite the small geographical scale (40 km) involved. Both mtDNA and microsatellites revealed a more pronounced differentiation between populations from two tributaries of a single river compared with their differentiation with a population from a neighbouring river. The comparison of wild and F₁-hatchery fish (produced from breeders originating from the same river) indicated significant changes in allele frequencies and losses of low-frequency alleles but no reduction in heterozygosity. Estimates of variance and inbreeding population size indicated that susceptibility to genetic drift and inbreeding in one population increased by twofold after only one generation of supplementation.     

Isozymes and DNA markers in gene conservation of forest trees

For long-lived plants that have to cope with high temporal and spatial environmental heterogeneity, genetic diversity is of prime importance for species persistence. Detrimental anthropogenic impact on the gene pool of forest trees calls for conservation of genetic resources. Potentials and limitations of isozymes and DNA markers in forest genetic conservation are reviewed. These markers can contribute to conservation with respect to the delimitations of species and hybrid zones, as well as the assessment of genetic diversity within and among populations. Markers are valuable to identify resource populations, since today‘s genetic diversity in forest trees is predominantly the result of plant history (e.g. glacial refuges, migration). Several suggestions have been put forward to optimize sampling of in situ or ex situ populations on the grounds of marker data. Restraint in this area is recommended. Different types of genetic markers (terpenes, isozymes, nuclear and extrachromosomal DNA polymorphisms) and quantitative traits yield different information about genetic diversity and population differentiation. Hence identification of resource populations should not solely be based upon a certain marker type or on quantitative traits alone. The capability of available markers to predict or assess adaptive potentials in forest tree populations is still very limited. This revised version was published online in August 2006 with corrections to the Cover Date.     

Uptake of Ga-67 into rat cerebral hemisphere and cerebellum. Comparison with Fe-59.

Transferrin and transferrin receptors play an important role in the transport of iron into the brain. To determine whether gallium enters the brain by the same mechanism, uptakes of Ga and 59Fe have been compared under controlled conditions. Rates of gallium penetration into brain (K) were four times slower than those for 59Fe. Kin for Ga when infused with citrate were 0.88 ± 0.24 and 0.94 ± 0.39 x 10 ml gh for cerebral hemisphere and cerebellum, respectively. When infused as the transferrin complex, Ga uptake into the brain was not different from that when infused with citrate. The presence of the anti-transferrin receptor antibody OX-26 significantly reduced uptake of Fe by 60% and 64% into cerebral hemisphere and cerebellum, respectively. By contrast, pretreatment of rats with OX-26 enhanced the uptake of Ga into brain, particularly when infused with citrate; mean increases in uptake of Ga were 120% and 144% for cerebral hemisphere and cerebellum, respectively. Purified Ga-transferrin was also taken up into both brain regions examined in the presence of OX-26. These results indicate that the transport of non-transferrin bound gallium is an important mechanism for gallium uptake into brain.     

甜味蛋白研究的新进展

甜味蛋白(thaumatin)是世界上已知最甜的物质,具有很大的应用前景.Thaumatin的基因核苷酸和蛋白质氨基酸序列都已测定.晶体分析表明它具有高稳定的四级结构.在味蕾小孔中发现了介导thaumatin发生作用的物质.Thaumatin自身的功能仍不清楚.在多种生物中发现了thaumatin类似蛋白质,具有不同的生物活性.用基因工程手段实现了thaumatin在多种原核和真核生物中的表达,但迄今仍未得到理想的基因工程产品.     

谷胱甘肽硫转移酶基因表达的调控

催化内源性或外源性亲电子化合物与谷胱甘肽(GSH)结合的谷胱甘肽硫转移酶(GST)超基因家族是一族解毒功能蛋白.其基因的表达通过不同的机制受多种物质的调控.根据最近文献资料,对调控谷胱甘肽硫转移酶基因表达的基因结构、调控机制及氧化应激对谷胱甘肽硫转移酶基因表达的调控作用等作一简要综述.     

EB病毒BNLF-1基因的分子生物学研究进展

位于EB病毒基因组U₅-TR区内的BNLF-1基因,其转译产物为潜伏膜蛋白(latent membrane protein1, LMP-1),由于LMP-1可以导致细胞转化并在EB病毒致癌过程中具有重要作用,因而成为近年来EB病毒分子生物学及相关肿瘤如人鼻咽癌、伯基特淋巴瘤、何杰金氏病等疾病病因发病学研究的热点,并取得了一批有重要意义的成果,文章从BNLF-1的基因结构及表达调控, LMP蛋白的结构及生化功能, LMP-1的生物学功能和LMP-1研究进行评述.