四个烙铁头蛇毒凝集素样蛋白基因的克隆与序列分析

从烙铁头蛇（Trimeresurus mucrosquamatus）的毒腺中提取mRNA，利用RT-PCR进行体外扩增，获得凝集素样蛋白基因，克隆至PMD18-T载体中，筛选出4种凝集素样蛋白基因（命名为TML-1、TML-2、TML-3和TML-4）。由基因序列推导出的氨基酸序列表明：TML-1，2，3，4序列中均有CRD结构。序列同源性比较和Cys位点分析推测：TML-1和TML-2可能分别是类似于flavocetin-A的蛇毒凝集素样蛋白的#alpha#亚基和#beta#亚基；TML-3可能类似于GPIb-bp的蛇毒凝集素样蛋白的#alpha#亚基，TML-4则可能是类似于IX/X-bp的蛇毒凝集素样蛋白的β亚基。

人、黑猩猩和叶猴FKN全基因的克隆及体外表达的比较研究

下载PDF阅读器目的:克隆人、黑猩猩和叶猴FKN全基因及体外表达,比较研究FKN在进化过程中基因组水平和蛋白表达水平的差异.方法:应用基因重叠延伸拼接PCR法(Gene splicing by overlapping extension PCR,SOE-PCR)将FKN的3个外显子编码序列依次进行前后拼接,然后插入pcDNA3.1/myc-His(-)A真核表达载体中,经酶切、测序鉴定后转染CHO细胞体外表达,RT-PCR、SDS-PAGE和Western blot检测其表达产物.结果:酶切、测序鉴定证实插入的基因片段为完整的FKN,RT-PCR可从转染的CHO细胞中扩增出一条与目的基因大小一致的DNA片段,其表达蛋白能分泌至胞外,SDS-PAGE显示其分子量约为95 000,抗c-myc抗体可与载体上的c-myc蛋白特异性结合.测序显示人、黑猩猩和叶猴相比,FKN基因除了有散在的点突变外,还发现有一明显的30 bp的缺失,但此缺失对FKN蛋白的表达并不影响.结论:成功克隆人、黑猩猩和叶猴FKN全基因,基因组水平和蛋白表达水平的比较研究为后续探讨FKN在高级灵长类物种进化过程中免疫学功能的演变奠定基础.

猕猴属五个种mtDNA多态性研究

结合日本猴(M. fuscata)的有关资, 料构建了猕猴属6个种的分子系统树, 并给出各个种的分化时间。这6个种可分成4个类群, 熊猴和藏酋猴、恒河猴和日本猴之间的遗传距离较近, 可分别划分为同一类群, 红面猴与其他5种猴的遗传距离最远, 在系统发生上分离最早。图9表5参29

除虫菊和茼蒿核糖体DNA ITS区的序列

中国科学院基金;其它部委、高等院校基金

单核苷酸多态性的研究技术

在对人类基因组单核苷酸多态性(SNP)的概念做简要说明的基础上,系统地介绍14种检测分析SNP的技术和方法的原理、操作要点及其优缺点。

洱海四种鲤鱼线粒体DNA遗传相似性的初步研究

用6种限制性内切酶对洱海的四种鲤鱼--洱海鲤(C. barbatus)、春鲤(C. longipectoralis)、大眼鲤(C. megalophthalmus)和杞麓鲤(C. carpio chila),其中前三种为洱海特有,进行了线粒体DNA(mtDNA)的限制性片段长度多态性(RFLP)分析,构建了它们的mtDNA限制性内切酶图谱。结果表明,这四种鲤鱼在种内和种间均缺乏mtDNA RFLP。这种现象在鱼类种间的mtDNA的RFLP研究中是罕见的。分析这一现象的原因,可能在于这些物种是同域形成物种,并且其分化时间还相当短。

青海湖裸鲤 mtDNA 遗传多样性的初步研究

该文用 BclⅠ、AvaⅠ、BamHⅠ、PstⅠ、KpnⅠ、PvuⅡ 共6种限制性内切核酸酶, 分析了15尾青海湖裸鲤 mtDNA 的限制性片段长度多态性, 共检测出20个酶切位点, 发现 BclⅠ、BamHⅠ和 PvuⅡ 三种酶切类型具有多态性. 根据不同个体 mtDNA 的酶切类型, 青海湖裸鲤存在4种 mtDNA 单倍型, 计算 mtDNA 多态度 π 值为 0.0043, 初步认为青海湖裸鲤在线粒体 DNA 上存在较丰富的群体内变异。

Drosophila auraria复合种Period基因Thr-Gly区段的初步研究

测定了金色果蝇复合种5个姐妹种和D. rufa的period (per)基因的Thr-Gly区段序列。该区段序列分析表明:DNA序列的碱基组成拥有果蝇其他基因的共同特点;颠换数多于转换数,两两物种间的颠换率与转换率的总比值为2-5,密码子第3位的颠换与转换的比值为2.5-5;同义替换/异义替换(Ks/Ka)值远大于10,且有的物种间根本不存在非同义突变,低的Ka值说明该复合种的per基因Thr-Gly区段在进化过程中可能承受着较强的选择压力。运用所得的核苷酸序列构建Drosophila auraria复合种的系统发育树,为澄清该类群的系统演化关系提供了新的线索。

细胞色素b基因序列与11种熊蜂的系统进化

通过比较一段357bp的细胞色素b的序列,分析了熊蜂属5亚属11种熊蜂系统发育关系。5亚属包括Bombus(5种)、Thoracobombus(3种)、Mendacibombus(1种)、Fervidobombus(1种)和Pyrobombus(1种)。该序列有65个单变异多态位点和71个简约信息多态位点,翻译成119个氨基酸序列后有45个氨基酸变异位点。根据P-距离构建的邻接树(NJ tree)和最大简约树(MP tree)都显示同样的结果:Mendacibombus(B. avinovielllus)分化最早;Fervidobombus(B. pensylvanicus)次之;Pyrobombus(B. impatiens)和Bombus形成姊妹群;Bombus亚属是单系群,其中B. ignitus在所研究的5个种中分化最早。

甘蔗细茎野生种云南不同生态类型的RAPD分析

云南甘蔗细茎野生种不同生态类型的遗传变异较大,具有丰富的遗传多样性;低纬度类型的遗传多样性明显高于高纬度类型,在相同的纬度范围内,随着海拔的升高,其多态性逐渐减少;基于分子聚类分析,86份材料被划分为8个不同群体,表现出明显的地理分布的特点。

PHYLOGENY OF RHESUS-MONKEYS (MACACA-MULATTA) AS REVEALED BY MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA, purified from 36 samples of 23 local populations which are widely distributed in Vietnam, Burma, and 10 provinces of China, has been analyzed to model the phylogeny of rhesus monkeys. The 20 local populations of China may represent nearly all major populations in China. Using 20 restriction endonucleases of 6-bp recognition, we observed a total of 50-61 sites in the various samples. By combining the cleavage patterns for each enzyme, the 36 samples were classified into 23 restriction types, each of which was found exclusively in the respective population from which samples were obtained By combining the earlier study of Indian rhesus monkeys, phylogenetic trees, which have been constructed on the basis of genetic distance, indicate that rhesus monkeys in China, Vietnam, India, and Burma can be divided into seven groups. Integrating morphological and geographical data, we suggest that rhesus monkeys in China, Vietnam, and Burma may be classified into six subspecies-M. m. mulatta, M. m. brevicaudus, M. m. lasiotis, M. m. littoralis, M. m. vestita, and M. m. tcheliensis-and rhesus monkeys in India may be another valid subspecies. M. m. tcheliensis is the most endangered subspecies in China. Divergence among subspecies may have begun 0.9-1.6 Ma. The radiation of rhesus monkeys in China may have spread from the southwest toward the east. The taxonomic status of the Hainan monkey and the Taiwan monkey require further investigation.

STUDIES ON THE CHROMOSOMES OF GENUS NYCTICEBUS

The karyotypes of three species (N. coucang, N. intermedius, and N. pygmaeus) of genus Nycticebus, collected from the southern Yunnan of China, have been studied. All individuals from three species possess 2n=50 chromosomes, and all chromosomes in their complement are biarm chromosomes. The karyotype of slow loris (N. coucang) is characterized by having a secondary constriction and Ag-NORs in the short arms of pair No. 1. The G-banding patterns of three species are very similar. Three species are found to have multiple Ag-NORs. In N. coucang, NORs were observed on five pairs (Nos. 1, 6, 9, 15, and 23) and in N. intermedius and N. pygmaeus, NORs were found on four pairs (Nos. 6, 9, 15, and 20). This finding indicates that slow lorises, as primitive primates, also have multiple NOR-bearing chromosomes. Finally, the classification of genus Nycticebus by karyotype analysis is discussed, and our results suggest that there are at least two valid species, namely: N. coucang and N. pygmaeus.

Adaptive diversification of bitter taste receptor genes in mammalian evolution

The diversity and evolution of bitter taste perception in mammals is not well understood. Recent discoveries of bitter taste receptor (T2R) genes provide an opportunity for a genetic approach to this question. We here report the identification of 10 and 30 putative T2R genes from the draft human and mouse genome sequences, respectively, in addition to the 23 and 6 previously known T2R genes from the two species. A phylogenetic analysis of the T2R genes suggests that they can be classified into three main groups, which are designated A, B, and C. Interestingly, while the one-to-one gene orthology between the human and mouse is common to group B and C genes, group A genes show a pattern of species- or lineage-specific duplication. It is possible that group B and C genes are necessary for detecting bitter tastants common to both humans and mice, whereas group A genes are used for species-specific bitter tastants. The analysis also reveals that phylogenetically closely related T2R genes are close in their chromosomal locations, demonstrating tandem gene duplication as the primary source of new T2Rs. For closely related paralogous genes, a rate of nonsynonymous nucleotide substitution significantly higher than the rate of synonymous substitution was observed in the extracellular regions of T2Rs, which are presumably involved in tastant-binding. This suggests the role of positive selection in the diversification of newly duplicated T2R genes. Because many natural poisonous substances are bitter, we conjecture that the mammalian T2R genes are under diversifying selection for the ability to recognize a diverse array of poisons that the organisms may encounter in exploring new habitats and diets.

Molecular phylogeny and biogeography of Oriental voles: genus Eothenomys (Muridae, Mammalia)

Oriental voles of the genus Eothenomys are predominantly distributed along the Southeastern shoulder of the Qinghai-Tibetan Plateau. Based on phylogenetic analyses of the mitochondrial cytochrome b gene (1143 bp) obtained from 23 specimens (eight species)