Serine/threonine protein phosphatases in DNA damage response

DNA damage response (DDR) is among the most important of the mechanisms that maintain genome stability which, when destabilized, predisposes organs to cancer. Reversible phosphorylation mediated by protein kinases and protein phosphatases regulates most, if not all, cellular activities, including DDR. Protein kinase inhibitors have become the main focus of targeted therapy and anticancer drug development. However, our limited knowledge of protein phosphatase function is compromising our capacity to develop therapeutic agents against phosphatases. In this review, we summarize the roles of serine/threonine protein phosphatases involved in DDR and propose that in situ dephosphorylation of phosphoproteins by protein phosphatases, instead of proteasome-mediated degradation of phosphoproteins, is mainly employed by cells.     

Mitochondrial DNA variation analysis suggests extreme low genetic diversity in Guizhou snub-nosed monkeys （Rhinopithecus brelichi）

Guizhou snub-nosed monkey (Rhinopithecus brelichi) is a unique, endangered primate in China, mainly distributed in Fanjing Mountain National Natural Reserve, Guizhou Province, in an area of 275 km2. Recently, habitat loss and fragmentation have caused population isolation. To assess genetic diversity within this species and its population structure, we sequenced 400 bp of the hypervariable I segment from the mitochondrial DNA control region for 128 individuals. Only one haplotype was identified from these individuals. Compared with other primate species, R. brelichi can be regarded as a species with very low genetic diversity, which further adds to the conservation concern.     

Rapid growth and sterility of growth hormone gene transgenic triploid carp

Triploid carp (100%) with 150 (3n=150) chromosomes were obtained by crossing the females of improved tetraploid hybrids (♀, 4n=200) of red crucian carp (♀)×common carp (♂) with the males of diploid yellow river carp (♂, 2n=100). The crosses yielded transgenic triploid carp (positive triploid fish, 44.2% of the progeny) and non-transgenic triploid carp (negative triploid fish). Histological examination of the gonads of 24-month-old positive triploid fish suggested they were sterile and the fish were not able to produce mature gametes during the breeding season. Morphologically, both the positive and negative triploid fish were similar. They had a spindle-shaped, laterally compressed, steel grey body with two pairs of barbells. Most of the quantifiable traits of the triploid carp were intermediate between those of the two parents. The positive and negative triploid fish were raised in the same pond for 2 years. The mean body weight of the positive triploid fish was 2.3 times higher than the negative triploid fish. The weight of the largest positive triploid fish was 2.91 times higher than that of the largest negative triploid fish. Thus, we produced fast-growing transgenic triploid carp that have a reduced ecological risk because of their inability to mate and produce progeny.     

Habitat-induced reciprocal transformation in the root phenotype of Oriental ginseng is associated with alteration in DNA methylation

Oriental ginseng is an important medicinal plant that grows in 2 major forms or ecotypes, wild and domesticated. Each form differs conspicuously in root phenotype, but can be converted from one type to another by habitat. Here we show that the habitat-induced transformation of ginseng root phenotype was accompanied by alteration in cytosine methylation at a large number of 5′-CCGG-3′ sites detected by the methylation-sensitive polymorphism (MSAP) marker. The collective CG and CHG methylation levels of all 4 landraces of the domesticated form were significantly lower than those of the wild form. Interestingly, artificially transplanted ginseng plants recreated in both directions the methylation levels (at least in CHG) of their natural counterparts. The methylation differences between the 2 ginseng ecotypes were validated at 2 isolated MSAP loci bearing homology to a 5S rRNA gene or a copia retrotransposon. Our results implicate a link between epigenetic variation and habitat-induced phenotypic flexibility in Oriental ginseng.     

不同倍性(2×、3×、4×)鸭梨基因组DNA甲基化水平与模式分析

以鸭梨(Pyrus.bretschneideri Rehd.cv.‘Yali')多倍体(2×、3×、4×)为材料,应用甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)技术研究不同倍性鸭梨基因组DNA甲基化水平及模式变化.共筛选22对引物,平均每对引物产生约50条带,统计100～500 bp的扩增带,共计954个位点.分析表明:鸭梨2×、3×、4×分别检测到831、886、852个位点,发生甲基化位点分别占15.5%、20.0%、16.8%,其中全甲基化占9.7%、11.9%、9.7%.分析这些位点甲基化模式表明:甲基化模式在三个倍性间类型相同的占所检测位点的69.2%,仅两种倍性相同的为25.4%,各不相同的占5.5%,其中三倍体41.4%甲基化位点发生改变,显示出独特的DNA甲基化特点.     

LAGE-1DNA肿瘤疫苗的抗肿瘤治疗效果研究

以LAGE-1转染EMT6构建小鼠乳腺癌肿瘤模型;将BalB/c小鼠随机分为4组,以pcDNA3.1/LAGE-1及对照pcDNA3.1分别接种实验组和对照组各三次.于末次免疫后10 d在小鼠左背部、右背部皮下分别种植EMT6肿瘤细胞和EMT6/LAGE1肿瘤细胞．荷瘤后观察成瘤时间、肿瘤大小,并对小鼠脾细胞进行CTL细胞杀伤活性实验来观察DNA疫苗引起的免疫反应.结果显示LAGE-1DNA疫苗在体内能诱导有效的特异性肿瘤免疫应答,该疫苗简便有效.     

利用环介导恒温扩增法简易诊断松萎蔫病Takuya Aikawa,Natsumi Kanzak,Taisei Kikuch

诊断松萎蔫病的传统方法是从木质组织中分离松材线虫,然后用显微镜进行形态学鉴定。基于DNA分子检测的方法尽管很灵敏,不需要固定发育阶段的松材线虫,但是仍然需要用Baermann 漏斗法分离线虫,并且需要昂贵的仪器。笔者研发了利用环介导恒温扩增法检测松材线虫的存在,该方法包括：(1) 从木块中提取DNA;(2) DNA 扩增;(3) 通过反应溶液的颜色做出诊断。该方法仅使用培养箱且整个过程只需要90 min,比以往方法更加简便和快速。     

用改进的酚-氯仿法提取榕小蜂基因组DNA及SRAP分析

以传粉榕小蜂为材料,用改进的酚-氯仿法对不同保存条件下的样品提取基因组DNA,并进行分光光度、琼脂糖凝胶电泳、SRAP扩增等方法的鉴定.结果表明:改进的酚-氯仿法均可从4种保存条件下的样品中提取出DNA,新鲜样品和体积分数95％乙醇固定样品提取效果比其他2组提取效果好.选择体积分数95％乙醇固定所提取的DNA为模板进行...     

剑尾鱼肾脏的超微结构?观察

应用光镜和透射电镜对剑尾鱼肾脏的组织学和超微结构进行了观察。结果表明:与其它硬骨鱼类相似,剑尾鱼中肾由肾小体和肾小管组成,肾小体分布较分散;肾小管分为颈段、近曲小管、远曲小管和集合管。肾脏中散布大量淋巴样组织,电镜水平上可以区分淋巴细胞、单核细胞和黑色素巨噬细胞等类型。本文还讨论了肾脏作为鱼类体内一个具有多种生理功能复合器官的结构特点。     

恐怖的“积木木乃伊’

2001年,考古学家宣布,在苏格兰史前村庄克拉德·哈兰遗址,出土了4具铜器时代居民的木乃伊（干尸）,它们分别属于一名3个月大的婴儿、一名年轻女性、一名40多岁的妇女和一名男性。然而,考古学家最近又宣布,这4具木乃伊其实是由不同家族成员的骨骸拼接而成的。研究发现,其中一位女性当初是依据骨盆特征来判定其性别的,而最新DNA...