草间钻头蛛(Hylyphantes graminicola)对果蝇(Drosophila melanogaster)捕食作用的研究

在实验室条件下,研究了草间钻头蛛Hylyphantes graminicola对果蝇Drosophila melanogaster的捕食功能反应.结果表明,在一定范围内,草间钻头蛛捕食效应随猎物密度增加而增加;随自身密度增加而减小;随着蜘蛛和果蝇密度的增加,相互干扰明显,捕食效率下降;雌蛛比雄蛛捕食量大.     

热效应在微生物诱变育种中的应用

本文综述了微生物诱变育种中的热效应机理及其应用研究进展。热在微生物诱变育种中具有热诱变效应和热筛选效应, 热诱变效应通过热引起DNA中G-C碱基对的置换实现, 热筛选效应可以从其他诱变剂诱变的菌体中获得更高的正向突变率。     

果蝇伴性隐性致死突变试验中芥子碱的辐射保护作用

过去的工作证明一些十字花科植物的抗辐射性与其体内存在的天然辐射保护物质有关。芥子碱是存在于十字花科植物中的一种天然辐射保护物质。它对大麦和小麦以及小鼠均有辐射保护作用。本文报道芥子碱在果蝇伴性隐性致死(SLRL)突变试验系统中的辐射保护作用。供试果蝇并不拒食芥子碱水溶液。10 mg/ml芥子碱对果蝇无生理毒性,对生育力亦无不良影响。40 Gy X 射线诱发8.96% SLRL 突变。而40 Gy 辐照前喂10 mg/ml芥子碱的果蝇其SLRL 突变率降为0.4%。Oregon K 果蝇的自发突变率为0-0.4%。可见喂芥子碱后辐射诱发的SLRL 突变不复出现,说明芥子碱对果蝇有很强的辐射保护作用。也讨论了芥子碱作为潜在抗癌物质的可能性。     

烟草水提物对果蝇寿命及有关基因转录的影响

探究烟草的水浸泡液对果蝇寿命与衰老情况的影响以及其中的分子机制。取羽化12 h内的新生W^1118系雄果蝇,分别培养于添加不同浓度烟草浸出液的培养基中处理,每日统计果蝇死亡情况;每7日提取对照组与最高浓度处理组果蝇全RNA,通过Real-time PCR检测过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、去泛素化酶(Rpn11)和去乙酰化酶(Sirt6)等与寿命有关基因的表达水平变化。结果显示,烟草影响使雄果蝇寿命显著缩短,且影响随烟草提取液浓度增加影响逐渐增大;烟草影响下果蝇的抗氧化基因出现显著上调,泛素化调节基因与去乙酰化基因等表达水平的变化均出现显著改变。表明烟草对果蝇的寿命情况有不利影响,可能通过引起氧化胁迫导致果蝇早衰。     

百日咳疫苗用菌株基因的遗传稳定性研究

目的对百日咳疫苗(pertussis vaccine)用菌株的主要保护性抗原基因序列的遗传稳定性进行研究,进一步验证此前一代测序的结果,为疫苗的安全性和有效性提供依据。方法采用第二代测序技术(next generation sequencing,NGS)对百日咳疫苗生产用菌株的主代代次(P4)、工作代次(P7)、疫苗代次(P13)及疫苗代次后连续传3代的第3代样品V3代次(P16)的主要抗原基因序列进行测序并分析。采用ELISA和Lowry法对百日咳疫苗生产用菌株各代次主要抗原组分进行抗原含量检测及比活性检测。结果百日咳毒素(pertussis toxin,PT) S1亚基上的8个等位基因氨基酸相对应的24个碱基位点中最高突变率为0.38%,其中疫苗代次最高突变位点(A2789C0.92%)在S5亚基上,其他各代次最高突变位点均不在S亚基上。PT全基因组最高突变率4%,且同一个位点不同代次的碱基突变呈不规律性,突变率没有随传代次数的增加而增高。百日咳黏附素(pertactin,PRN)的24个抗原表位中最高突变率在疫苗代次T951G位点,突变率为1.66%;其他位点的突变率均0.8%。全基因组中突变率最高的位点为V3代次C2025T,突变率为6.24%;各代次PRN全基因组最高突变率6.24%,且不在主要抗原表位区域内;在同一个位点不同代次的碱基突变没有随传代次数的增加而增高,呈不规律性。丝状血凝素(filamentous hemagglutinin,FHA)中B细胞抗原表位区中碱基最高突变率为0.98%(V3代次T6449G位点),其他代次均0.8%;全基因组各代次位点最高突变率为V3代次C7323G,突变率为9.89%;其余代次突变率均9.89%,且不在抗原表位区,同一个位点不同代次的碱基突变没有随传代次数的增加而增高,呈不规律性。PT的ELISA比活性平均值为0.532±0.100,最低限值为0.232。PRN的ELISA比活性平均值为0.885±0.110,最低限值为0.555。FHA的ELISA比活性平均值为0.701±0.170,最低限值为0.191,百日咳菌种抗原含量和比活性较稳定。结论传代过程中PT、PRN、FHA主要碱基位点突变率较低,说明百日咳菌种基因遗传稳定性好。     

绿色木霉纤维素酶高产突变型的诱变和筛选

    1．通过各种化学和物理因素的复合处理，诱发T. vi,ide 2” 的变异，得到9株较好的突变型。其中典型的是绿色突变型403-2，其纤维素酶活较高、稳定，平均CMC酶活为101毫克／毫升，滤纸酶活为9.3毫克／毫升，比亲本菌株提高3倍。    2．在所使用的诱变剂中以紫外光和亚硝酸为好，特别是将它们进行复合处理时效果更佳。诱变时，降低诱变剂的相对剂量,即缩短处理时间，正变率大幅度提高。    3.诱变之前采用限量培养液使抱子萌动，可以降低相对剂量，增加抱子对诱变剂的敏感性，提高突变率，增加正变效果。    4.经诱变后得到的突变型遗传性不易稳定，往往发生退化。试验证明，用连续优势选择方法可以选育出种性纯、产量高而稳定的突变型。     5. C 酶是降解夭然纤维素必不可少的成分，因而，用C,酶活表示纤维素酶的离低，菌种的好坏是合理的。测C,酶的底物最好是滤纸。     

科技文摘

黄色猩猩蝇是一种体长1厘米、体重1毫克的小果蝇,是遗传学研究中常用的材料。这果蝇中有一系统因诱变剂的作用而失去了飞行能力,其中一种是因为不能合成肌动蛋白(一种肌肉蛋白)。东京大学理学部物理学教研室的堀田凯树副教授等给这种果蝇的受精卵注入肌动蛋白基因,成功地使果蝇重新飞了起来。他们将肌动蛋白基因同称作转座子的跳动基因连接起来,然后以40万倍的基因量将它注入果蝇的受精卵。结果该受精卵发育成幼蝇后有30％恢复了飞行能力。肌动蛋白基因进入生殖细胞的染色体中,以后遗传给子代,这个过程就是通常所说的“基因治疗”。将基因注入受精卵,该基因的功能在其子代中出     

果蝇视觉学习记忆能力与脑部cAMP水平的相关性

利用闭环飞行模拟系统研究果蝇视觉飞行定向行为的操作式条件化 ,证明正常果蝇视觉学习记忆能力与日龄有关 ,即 3～ 4d龄果蝇的学习记忆能力明显优于1～ 2d龄果蝇 ,蝇脑内的cAMP含量也呈现随果蝇日龄增加而增加的趋势 .同时对学习记忆缺陷型果蝇进行检测 ,其脑内cAMP含量高于正常对照组果蝇 .通过喂食PDEase抑制剂咖啡因扰乱cAMP代谢 ,使果蝇cAMP水平异常提高 ,导致果蝇学习记忆能力显著下降 ,表明果蝇视觉学习记忆需要脑内cAMP水平处于一适当范围 ,过高或过低的cAMP水平都将影响果蝇的视觉学习记忆能力     

自由基、营养、天然抗氧化剂与衰老

影响衰老的因素很多,其中主要有遗传基因、饮食营养、生活方式、运动多少、心理状态、医疗条件及环境因素等。其中饮食营养是非常重要的因素,而且是可以控制的因素。研究证明,营养不良和营养过剩都会影响健康和寿命,而健康和延长寿命最有效的方法是限食(限能)。最近,美国和英国的权威杂志《Science》和《Nature》发表研究文章表明,节食,以及一些自由基清除剂,如小分子多酚类物质白藜芦醇,可以启动长寿基因SIRI1,抑制肿瘤基因p53,阻断细胞凋亡,延缓衰老和延长寿命。早在1955年,Dr. Harman发表的“衰老的自由基理论”就提出,体内产生过多自由基是引起衰老的重要因素,保持体内自由基和抗氧化剂的平衡可以延缓衰老。我们的实验还证明,天然抗氧化剂茶多酚可延长果蝇的寿命,使果蝇匀浆中的超氧化物歧化酶(superoxide dismutase,SOD)活性增加,脂质过氧化水平降低,改善高脂引起的果蝇寿命缩短、SOD活性降低以及脂质过氧化水平的升高。作者最近的实验还证明,茶多酚可以防止氧化应激引起的线虫寿命的缩短,还可以预防和治疗6-OHDA引起的大鼠帕金森氏综合症,山楂黄酮可以预防和治疗蒙古沙鼠缺血引起的中风,大豆异黄酮和尼古丁可以预防和治疗转基因线虫和小鼠老年痴呆症。饮食营养和天然抗氧化剂研究的进展将对人类健康和延缓衰老,提供新的线索并做出重大贡献。     

抗氧化剂对脂质过氧化反应混合物引起人培养细胞DNA损伤的影响

 <正> 脂质过氧化反应(LPR)中产生的自由基和过氧化物可引起多种细胞损伤,特别是对DNA的损伤可能是许多癌肿的重要原因。Akasaka等将E.coli与鼠肝微粒体脂质过氧化反应混合物(LPRM)孵育,使有DNA修复缺陷的细菌株突变率增加。我们将人羊膜FL细胞与大鼠肝微粒体LPRM共同培养,看能否诱发程序外DNA合成(UDS),并分组加入抗氧化剂观察其影响。     

Why is the genome variable? Insights from Drosophila.

The analysis of variation in DNA restriction maps and DNA sequence in natural populations of Drosophila melanogaster and related species has revealed a remarkable richness of diversity. This review describes some of the results of population genetic studies of this variation that are beginning to reveal how interactions between natural selection, genetic drift, mutation rate, recombination rate and population size have contributed to the observed patterns.     

High density of long dinucleotide microsatellites in Drosophila subobscura

We isolated 96 dinucleotide repeats with five or more tandemly repeated units from a subgenomic Drosophila subobscura library. The mean repeat unit length of microsatellite clones in D. subobscura is 15, higher than that observed in other Drosophila species. Population variation was assayed in 32-40 chromosomes from Barcelona, Spain, using 18 randomly chosen microsatellite loci. Positive correlation between measures of variation and perfect repeat length measures (mean size, most common, and longest allele) is consistent with a higher mutation rate in loci with longer repeat units. Levels of microsatellite variation measured as variance in repeat number and heterozygosity in D. subobscura were similar to those of Drosophila pseudoobscura and higher than those of Drosophila melanogaster and Drosophila simulans. Our data suggest that higher levels of microsatellite variation, and possibly density, in D. subobscura compared with D. melanogaster are due to both a higher average effective population and a higher intrinsic slippage rate in the former species.     

The spontaneous mutation rate in Drosophila simulans

The purpose of this investigation was to determine whether there exists a correlation in Drosophila between the spontaneous mutation rate and the amount of dispersed middle repetitive (mobile) DNA sequences. The amount of these sequences is 7 times less in Drosophila simulans as compared to Drosophila melanogaster. Therefore, if a correlation exists, the spontaneous mutation rate in Drosophila simulans should be 7 times lower than that in Drosophila melanogaster. We isolated an X-chromosome inversion after X-irradiation of wild-type Drosophila simulans males, that reduced crossing-over between white and forked, two X-linked visible markers, to less than 1%. This inversion was subsequently used to determine the sex-linked recessive lethal mutation rate in Drosophila simulans males of a laboratory strain marked with white. The frequency of these lethal mutations found is not different from that observed in Drosophila melanogaster males of laboratory strains.     

Deleterious Background Selection with Recombination

An analytic expression for the expected nucleotide diversity is obtained for a neutral locus in a region with deleterious mutation and recombination. Our analytic results are used to predict levels of variation for the entire third chromosome of Drosophila melanogaster. The predictions are consistent with the low levels of variation that have been observed at loci near the centromeres of the third chromosome of D. melanogaster. However, the low levels of variation observed near the tips of this chromosome are not predicted using currently available estimates of the deleterious mutation rate and of selection coefficients. If considerably smaller selection coefficients are assumed, the low observed levels of variation at the tips of the third chromosome are consistent with the background selection model.     

The coalescent process in models with selection, recombination and geographic subdivision

A population genetic model with a single locus at which balancing selection acts and many linked loci at which neutral mutations can occur is analysed using the coalescent approach. The model incorporates geographic subdivision with migration, as well as mutation, recombination, and genetic drift of neutral variation. It is found that geographic subdivision can affect genetic variation even with high rates of migration, providing that selection is strong enough to maintain different allele frequencies at the selected locus. Published sequence data from the alcohol dehydrogenase locus of Drosophila melanogaster are found to fit the proposed model slightly better than a similar model without subdivision.     

A method for estimating the mutation,gene conversion and recombination parameters in small multigene families

A simple two-locus gene conversion model is considered to investigate the amounts of DNA variation and linkage disequilibrium in small multigene families. The exact solutions for the expectations and variances of the amounts of variation within and between two loci are obtained. It is shown that gene conversion increases the amount of variation within each locus and decreases the amount of variation between two loci. The expectation and variance of the amount of linkage disequilibrium are also obtained. Gene conversion generates positive linkage disequilibrium and the degree of linkage disequilibrium decreases as the recombination rate is increased. Using the theoretical results, a method for estimating the mutation, gene conversion, and recombination parameters is developed and applied to the data of the Amy multigene family in Drosophila melanogaster. The gene conversion rate is estimated to be approximately 60-165 times higher than the mutation rate for synonymous sites.     

The mutation rates of di-, tri- and tetranucleotide repeats in Drosophila melanogaster

In a recent study, we reported that the combined average mutation rate of 10 di-, 6 tri-, and 8 tetranucleotide repeats in Drosophila melanogaster was 6.3 x 10(-6) mutations per locus per generation, a rate substantially below that of microsatellite repeat units in mammals studied to date (range = 10(-2)-10(-5) per locus per generation). To obtain a more precise estimate of mutation rate for dinucleotide repeat motifs alone, we assayed 39 new dinucleotide repeat microsatellite loci in the mutation accumulation lines from our earlier study. Our estimate of mutation rate for a total of 49 dinucleotide repeats is 9.3 x 10(-6) per locus per generation, only slightly higher than the estimate from our earlier study. We also estimated the relative difference in microsatellite mutation rate among di-, tri-, and tetranucleotide repeats in the genome of D. melanogaster using a method based on population variation, and we found that tri- and tetranucleotide repeats mutate at rates 6.4 and 8.4 times slower than that of dinucleotide repeats, respectively. The slower mutation rates of tri- and tetranucleotide repeats appear to be associated with a relatively short repeat unit length of these repeat motifs in the genome of D. melanogaster. A positive correlation between repeat unit length and allelic variation suggests that mutation rate increases as the repeat unit lengths of microsatellites increase.      

High mutation rate of a long microsatellite allele in Drosophila melanogaster provides evidence for allele-specific mutation rates

Within recent years, microsatellite have become one of the most powerful genetic markers in biology. For several mammalian species, microsatellite mutation rates have been estimated on the order of 10(- 3)-10(-5). A recent study, however, demonstrated mutation rates in Drosophila melanogaster of at least one order of magnitude lower than those in mammals. To further test this result, we examined mutation rates of different microsatellite loci using a larger sample size. We screened 24 microsatellite loci in 119 D. melanogaster lines maintained for approximately 250 generations and detected 9 microsatellite mutations. The average mutation rate of 6.3 x 10(-6) is identical to the mutation rate from a previous study. Most interestingly, all nine mutations occurred at the same allele of one locus (DROYANETSB). This hypermutable allele has 28 dinucleotide repeats and is among the longest microsatellite reported in D. melanogaster. The allele-specific mutation rate of 3.0 x 10(-4) per generation is within the range of mammalian mutation rates. Future microsatellite analyses will have to account for the dramatic differences in allele-specific mutation rates.      

Low-level chemiluminescence from Drosophila melanogaster fed with chemical mutagens polycyclic aromatic hydrocarbon quinones and a carcinogenic bracken fern

The spontaneous photon emission (chemiluminescence) from Drosophila melanogaster fed chemical mutagens, polycyclic aromatic hydrocarbon quinones, and a carcinogenic bracken fern was studied. The fly chemiluminescence was evidently enhanced by mutagen or carcinogen administration and was increased proportionally to the administered amount of tested compound. Strong chemiluminescence was observed especially at the larval stage. Living larvae emitted stronger chemiluminescence than their homogenate. The chemiluminescence from Drosophila melanogaster fed polycyclic aromatic hydrocarbon quinones showed a linear relation with the mutation frequency in the Drosophila wing spot test. The chemiluminescence from flies fed a bracken fern decreased by the addition of free radical scavengers and active oxygen quenchers. The phosphatidylcholine hydroperoxide concentration in the flies was increased proportionally with the chemiluminescence intensity. It seems that the free radical formation is stimulated as shown by the enhanced chemiluminescence in mutagen- or carcinogen-dosed flies, and as a result, lipid peroxide accumulation accompanies mutation in Drosophila melanogaster.     

Inferring parameters of mutation, selection and demography from patterns of synonymous site evolution in Drosophila

Selection acting on codon usage can cause patterns of synonymous evolution to deviate considerably from those expected under neutrality. To investigate the quantitative relationship between parameters of mutation, selection, and demography, and patterns of synonymous site divergence, we have developed a novel combination of population genetic models and likelihood methods of phylogenetic sequence analysis. Comparing 50 orthologous gene pairs from Drosophila melanogaster and D. virilis and 27 from D. melanogaster and D. simulans, we show considerable variation between amino acids and genes in the strength of selection acting on codon usage and find evidence for both long-term and short-term changes in the strength of selection between species. Remarkably, D. melanogaster shows no evidence of current selection on codon usage, while its sister species D. simulans experiences only half the selection pressure for codon usage of their common ancestor. We also find evidence for considerable base asymmetries in the rate of mutation, such that the average synonymous mutation rate is 20-30% higher than in noncoding regions. A Bayesian approach is adopted to investigate how accounting for selection on codon usage influences estimates of the parameters of mutation.