An efficient method for multiple site-directed mutagenesis using type IIs restriction enzymes

Site-directed mutagenesis (SDM) methods are very important in modern molecular biology, biochemistry, and protein engineering. Here, we present a novel SDM method that can be used for multiple mutation generation using type IIs restriction enzymes. This approach is faster and more convenient than the overlap polymerase chain reaction (PCR) method due to its having fewer reaction steps and being cheaper than, but as convenient as, enzymatic assembly. We illustrate the usefulness of our method by introducing three mutations into the bacterial Streptococcus thermophilus Cas9 (bStCas9) gene, converting the humanized S. thermophilus Cas9 (hStCas9) gene into nuclease dead or H847A nickase mutants and generating sunnyTALEN mutagenesis from a wild-type TALEN backbone.     

栖息地特征对褐马鸡种群密度和集群行为的影响

1998-2000年冬季在山西芦芽山自然保护区选择车道沟和梅洞 2个研究地,就栖息地特征对褐马鸡 (Cros soptilonmantchuricum)越冬种群密度和集群大小的影响进行了研究。经χ2 检验,针叶林是褐马鸡冬季经常利用的栖息地类型。对 2个研究地的栖息地结构比较发现,车道沟适宜栖息地的面积较梅洞大,连接性也较好,褐马鸡种群有更为广阔的取食空间和更多的隐蔽场所,而且活动的阻碍也较小;而梅洞栖息地的取食空间较小,隐蔽场所较少,其活动时受到的阻碍也较大。从 2个研究地微生境结构特征的差异,并结合褐马鸡越冬期栖息地选择来看,树高和高层盖度对褐马鸡在 2个栖息地的选择上起重要作用。车道沟乔木较粗大,不但有丰富的食物,而且可提供较好的夜宿条件;高层盖度越大,栖息地的隐蔽条件越好,能吸引更多的褐马鸡个体取食和越冬。研究结果表明,栖息地结构连接性、微生境结构特征的差异以及人为干扰是褐马鸡种群密度和集群行为差异的主要影响因素。     

秦岭羚牛的采食行为

从集群、迁移、时间、方式、饮水和舔盐 6个方面报道了秦岭羚牛的采食行为。羚牛主要以集群的方式在白天采食 ,也见到羚牛单独采食及在夜间活动采食的情形。研究期间集群活动的羚牛个体数占所见羚牛总数的 95 4% ,有 5 0 %以上的羚牛是在大于 1 5只的羚牛群体中见到的。头牛在羚牛群体的采食迁移过程中起着重要的作用。它通过发出低沉的吼叫声来使牛群聚集在一起并共同采食迁移。随着食物的季节性生长变化 ,羚牛具有季节性上下垂直迁移采食的习性。羚牛采食时 ,多用上下唇扯断植物的枝 (茎 )叶 ,而不是以舌卷食 ,也用牙咬或啃食木本植物的幼枝或皮。羚牛有多种采食方式 ,一般是以常规行走的方式采食。但当食物超出正常采食方式所能获得的特殊情况下 ,羚牛会采用一些特殊的方式取食 ,包括后肢站立采食、骑树采食、压枝采食、撞击采食和跪地采食。     

长爪沙鼠(Meriones unguiculatus)和金黄地鼠(Mesocricetus auratus)的肺皮蒸发失水量研究

通过对长爪沙鼠和金黄地鼠肺皮蒸发失水量的研究表明,同种内个体间肺皮蒸发失水量与动物体重呈负指数相关。在10-30℃范围内,肺皮蒸发失水量随温度上升呈指数式增加。随着相对湿度的增加,肺皮蒸发失水量呈指数式减少。在10℃和20℃环境温度下,金黄地鼠肺皮蒸发失水量略高于长爪沙鼠,在30℃环境温度下,长爪沙鼠肺皮蒸发失水量略高于金黄地鼠。     

笼养环颈雉性选择的研究

环颈雉求偶炫耀有四种常见类型,即点头炫耀,侧炫耀,振翅鸣叫和追逐,各种类型求偶活动的强度存在显著的个体差异,剪尾和缩短距长都可使雄鸟的求偶频率下降,雌雉选择配偶受雄鸟所在笼舍条件及雄鸟某些身体特征的影响,条件较好的笼舍中的雄鸟往往具有较多的配偶,剪短雉鸟的尾羽对雌鸟的配偶选择无明显影响,而缩短距长则对雌鸟的择偶产生十分显著的影响,导致雄鸟配偶数的大幅度减少。     

秦岭羚牛群体分离与重组的变化

对4只佩戴无线电颈圈的羚牛进行直接观察的结果表明,羚牛群体的结构和组成是经常发生变化的。在羚牛各集群类型中,家群的稳定性最高,社群次之,混合群较差。虽然家群具有较高的稳定性,但是除母牛及其幼仔外,家群的其它成员也经常变化,羚牛的亚成体也不是构成家群的稳定成员。繁殖是羚牛集群类型变化的主要原因之一。在繁殖期中健壮的成年雄性个体因寻找配偶离开原群,在各群间游荡,成为羚牛群中最不稳定的成员。亚成体在其母亲产下幼仔后可能会中断与母牛间的关系,进入混合群和社群。羚牛反捕食策略和护幼行为也会影响羚牛群的分群和重组。     

秦岭羚牛的集群类型

1995 年7～11月,1996 年3～8月在陕西佛坪国家级自然保护区用野外直接观察和无线电跟踪器追踪的方法, 对秦岭羚牛( Budorcas taxicolor bedfordi) 的集群类型进行了观察。工作期间共记录到羚牛146 群次, 按群体大小和结构可将羚牛群分为家群、混合群、社群、聚集群和独牛5 种类型。在野外记录到的家群共32群次, 混合群15次, 社群47次, 聚集群2次, 独牛50 次。家群的成员是具有亲缘关系的个体, 在羚牛各类型的集群中占据核心的地位,是构成其它类型的羚牛群的基本单位。混合群的成员或是以家群为核心混杂以多只亚成体组成、或全部由亚成体和性成熟不久的年轻成体组成, 是介于家群和社群之间的一种集群方式; 其形成与羚牛幼仔在成长过程中社会习性的形成和完善有关。社群是由多个家群构成的相对比较稳定的一种群体, 在各季节中出现的比例均很稳定, 是羚牛群栖的主要形式。聚集群仅见于夏季, 是临时聚集的一种不稳定的集群类型, 其形成与采食和繁殖活动有关。独牛是羚牛集群的一种特殊形式, 繁殖季节出现的频率高于非繁殖季节。     

Genetic population structure in Siniperca scherzeri (Perciformes: Siniperca) in China inferred from mitochondrial DNA sequences and microsatellite loci

This study aims to improve the estimates of fine-scale genetic diversity and to compare the population structure of Siniperca scherzeri with nuclear and mitochondrial DNA (mtDNA) markers. We first combined mtDNA sequences and seven microsatellite loci to examine the genetic diversity of S. scherzeri in China. Phylogenetic and nested clade analyses revealed two major mtDNA lineages and four subclades. Bayesian multilocus genotype clustering from the microsatellite loci revealed that regional divergence estimates were quantitatively congruent between marker classes. These lineages should be recognized as the basic evolutionary significant units for S. scherzeri in China. Contradict to the previous studies on Coreoperca whiteheadi and Siniperca chuatsi, neither the Nanling–Wuyi Mountain range nor the Qingling–Dabie Mountain range represented a major phylogeographic barrier for S. scherzeri. Our fine-scale analyses of the genetic population structure of S. scherzeri provided insights into the evolutionary processes that shaped the genetic heterogeneity of this fish. This study may serve as a basis for the protection, monitoring and breeding improvement of germplasm resources of this commercially important fish.     

Study on the Interaction Between Nucleic Acids and Acetamiprid

Abstract

The interaction between deoxyribonucleic acid (DNA) and acetamiprid was studied. It was found that the fluorescence of acetamiprid could be enhanced in the presence of DNA in sulfuric acid solution. The excitation and emission wavelength of acetamiprid was 291 nm and 587 nm, respectively. Under optimal conditions, the calibration graph is over the range of 0.1–10 µg mL^?1. The calibration limit is 0.06 µg mL^?1 (S/N = 3). The determination results of DNA in yeast cell and golden staphylococcus samples by this method were satisfactory. The mechanism of the reaction is discussed.