水稻Phi类谷胱甘肽转移酶基因家族的功能分化研究

在植物中大多数功能基因是以基因家族的形式存在的，而基因重复则是基因家族的一种重要的进化方式。很多基因往往是由重复事件产生形成不同的拷贝，进而分化形成基因家族。谷胱甘肽转移酶（GSTs）是一类古老、庞大、行使解毒、抗逆、信号转导等多种功能的一个基因家族。本研究以栽培水稻（Oryza sativa ssp. japonica c.v. Nipponbare）为研究材料，以栽培水稻的Phi类GST的5个基因（OsGSTF3、OsGSTF6、OsGSTF14、OsGSTF15、OsGSTF16）为研究对象，分析了它们的系统发生和起源历史、不同组织的差异性表达、编码蛋白质的功能差异等问题，探讨了基因重复后5个基因的功能变化，主要结果如下： 1. OsGSTF3、OsGSTF14、OsGSTF15、OsGSTF16由串联重复产生，而OsGSTF6则由DNA转座产生;它们起源时间早在稻属（Oryza）分化之前。 2. 对水稻不同部位组织的RT-PCR结果表明这5个基因在水稻中的特异性表达组织部位有较大差异：OsGSTF3基因在叶、叶鞘、茎、根4个部位均有大量表达;OsGSTF6基因仅在叶中有表达;OsGSTF14基因在叶鞘、茎2个部位中有表达;OsGSTF15基因在茎、根2个部位中有表达;OsGSTF16则在叶、茎、根3个部位中有表达。 3. 将这5个基因连接原核表达载体PET30a并转化大肠杆菌BL21（DE3），获得了高表达菌株。将表达菌株进行大量表达，表达形式分析显示OsGSTF3蛋白是可溶性表达，而其余4个蛋白以包涵体的形式表达。通过亲和层析获得了纯化的OsGSTF3融合蛋白，OsGSTF3融合蛋白对底物CDNB和NBD-Cl具有高活性，酶动力学分析显示OsGSTF3融合蛋白对GSH与NBD-Cl有较高的亲和力，热力学分析显示该蛋白在40℃以下是热稳定的。通过对包涵体进行洗涤、亲和层析获得了纯化的OsGSTF6、OsGSTF14、OsGSTF15、OsGSTF16的融合蛋白，OsGSTF14融合蛋白对NBD-Cl有微弱活性，OsGSTF15融合蛋白对NBC有较高的活性，而没有检测到OsGSTF6与OsGSTF16融合蛋白的活性。

17 beta-Estradiol and progesterone improve in-vitro cytoplasmic maturation of oocytes from unstimulated prepubertal and adult rhesus monkeys

BACKGROUND: Effects of 17beta-estradiol and progesterone on rhesus monkey oocyte maturation in vitro were evaluated by embryo development subsequent to IVF. METHODS AND RESULTS: In experiment 1, immature cumulus-oocyte complexes collected from unstimulated adult females during the non-breeding season were matured in modified medium CMRL-1066 containing various combinations of gonadotrophins (FSH + LH), estradiol and/or progesterone. Formation of morulae and blastocysts was greatest in oocytes matured in medium containing estradiol and/or progesterone, with or without gonadotrophins (morula 38-46%, blastocyst 14-20%) than in control oocytes matured without estradiol or progesterone (morula 14%, blastocyst 0%). In experiment 2, cumulus-oocyte complexes from unstimulated prepubertal female monkeys were matured in medium with gonadotrophins, estradiol or progesterone. The best development to the morula stage was obtained with oocytes matured with gonadotrophins and estradiol or gonadotrophins and progesterone (43 and 25 morulae, respectively), while control oocytes matured with gonadotrophins but without steroid hormones gave the poorest morula developmental response (12%). However, there was no difference in blastocyst development across all groups (0-3%). CONCLUSIONS: These results demonstrate that during rhesus monkey oocyte maturation in vitro: (i) estradiol or progesterone can improve oocyte developmental competence; (ii) immature oocytes from prepubertal versus adult females have differential responses to challenge with estradiol or progesterone.

beta gamma-CAT, a non-lens beta gamma-crystallin and trefoil factor complex from amphibian skin secretions, caused endothelium-dependent myocardial depression in isolated rabbit hearts

Previous in vivo study demonstrated that beta gamma-CAT, a newly identified non-lens beta gamma-crystallin and trefoil factor complex from frog Bombina maxima skin secretions, possessed potent lethal toxicity on mammals resulted from hypotension and cardi

Melanoma cell growth inhibition by beta gamma-CAT, which is a novel non-lens betagamma-crystallin and trefoil factor complex from frog Bombina maxima skin

In vertebrates, non-lens beta gamma-crystallins are widely expressed in various tissues but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained

Acute toxicity of beta gamma-CAT, a naturally existing non-lens beta gamma-crystallin and trefoil factor complex from frog Bombina maxima skin secretions

In vertebrates, non-lens beta gamma-crystallins are widely expressed in various tissues, but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained

Phylogenetic studies of pantherine cats (Felidae) based on multiple genes, with novel application of nuclear beta-fibrinogen intron 7 to carnivores

The pantherine lineage of the cat family Felidae (order: Carnivora) includes five big cats of genus Panthera and a great many midsized cats known worldwide. Presumably because of their recent and rapid radiation, the evolutionary relationship among pantherines remains ambiguous. We provide an independent assessment of the evolutionary history of pantherine lineage using two complete mitochondrial (mt) genes (ND2 and ND4) and the nuclear beta-fibrmogen intron 7 gene, whose utility in carnivoran phylogeny was first explored. The available four mt (ND5, cytb, 12S, and 16SrRNA) and two nuclear (IRBP and TTR) sequence loci were also combined to reconstruct phylogeny of 14 closely related cat species. Our analyses of combined mt data (six genes; approximate to 3750 bp) and combined mt and nuclear data (nine genes; approximate to 6500 bp) obtained identical tree topologies, which were well-resolved and strongly supported for almost all nodes. Monophyly of Panthera genus in pantherine lineage was confirmed and interspecific affinities within this genus revealed a novel branching pattern, with P. tigris diverging first in Panthera genus, followed by P. onca, P. leo, and last two sister species P. pardus and P. uncia. In addition, close association of Neofelis nebulosa to Panthera, the phylogenetic redefinition of Otocolobus manil within the domestic cat group, and the relatedness of Acinonyx jubatus and Puma concolor were all important findings in the resulting phylogenies. The potential utilities of nine different genes for phylogenetic resolution of closely related pantherine species were also evaluated, with special interest in that of the novel nuclear beta-fibrinogen intron 7. (c) 2005 Elsevier Inc. All rights reserved.

New Insights into the Evolution of Intronic Sequences of the beta-fibrinogen Gene and Their Application in Reconstructing Mustelid Phylogeny

Mustelidae is the largest and most diverse family in the order Carnivora. The phylogenetic relationships among the subfamilies have especially long been a focus of study. Herein we are among the first to employ two new introns (4 and 7) of the nuclear P-f

Evidence of gene conversion in the evolutionary process of the codon 41/42 (-CTTT) mutation causing beta-thalassemia in southern China

The 4-bp deletion (-CTTT) at codon 41/42 (CD41/42) of the human beta-globin gene represents one of the most common beta-thalassemia mutations in East Asia and Southeast Asia, which is historically afflicted with endemic malaria, thus hypothetically evolvi

Aniracetam restores the effects of amyloid-beta protein or ageing on membrane fluidity and intracellular calcium concentration in mice synaptosomes

In the present study, we observed the in vitro effect of aniracetam on membrane fluidity and free calcium concentrations (Ca(2+)i) of frontal cortical (FC) and hippocampal (HP) synaptosomes of aged mice and young mice treated with amyloid-beta protein (A

A beta-galactose-specific lectin isolated from the marine worm Chaetopterus variopedatus possesses anti-HIV-1 activity

A 30 kDa beta-galactose-specific lectin named CVL was isolated from the polychaete marine worm Chaetopterus variopedatus (Annelida) and its anti-HIV-1 activity in vitro was determined. Results showed that CVL inhibited cytopathic effect induced by HIV-1 a

Synthesis and biological evaluation of N-acetyl-beta-aryl-1,2-didehydroethylamines as new HIV-1 RT inhibitors in vitro

A variety of N-acetyl-o-aryl-1,2-didehydroethylamines were synthesized by direct reduction-acetylation of beta-aryl-nitroolefins and assayed as HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) for the first time. Compound 7a exhibited a TI v