Ester production in E. coli and C. acetobutylicum

Genetic engineering has improved the product yield of a variety of compounds by overexpressing, inactivating, or introducing new genes in microbial systems. The production of flavor-enhancing ester compounds is an emerging area of heterologous gene expression for desired product yield in Escherichia coli. Isoamyl acetate, butyl acetate, ethyl acetate, and butyl butyrate are reported here to be produced by expressing Saccharomyces cerevisiae genes ATF1 or ATF2 and the strawberry gene SAAT in E. coli when the appropriate substrates are provided. Increasing the concentration of alcohol added to the reaction generally resulted in increased ester production. ATF1 expression was found to produce more isoamyl acetate and butyl acetate than ATF2 expression or SAAT expression in the strains and culture conditions examined. Additionally, SAAT expression resulted in greater isoamyl acetate and butyl acetate production than ATF2 expression. Butyl butyrate is produced by cell-free extracts of E. coli harboring SAAT but not ATF1 or ATF2.     

Mutagenic DNA repair in Escherichia coli, XX. Overproduction of UmuD'' protein results in suppression of the umuC36 mutation in excision defective bacteria

Overproduction of Umu⁺ or UmuD′ protein by means of a gene carried on a multicopy plasmid suppressed the umuC36 phenotype and permitted induction of mutations by ultraviolet light. The umuC122::Tn5 phenotype was not suppressed. Suppression of the umuC36 phenotype was only seen when excision repair was blocked by acriflavine or by an uvrA or uvrB mutation. Cleavage of UmuD to UmuD′ in SOS-induced cells was not dependent upon the presence of UmuC protein. The results are interpreted in terms of a revised model in which UmuC protein is envisaged as guiding UmuD′ to ReA protein which has recognized and become bound to an appropriate DNA lesion. It is suggested that the umuC36 mutation gives rise to a protein with reduced affinity for UmuD′ and that the effect of this can be compensated by an excess of UmuD′.     

仰鼻猴属(RHINOPITHECUS)的系统分类地位

自Milne-Edwards(1868-1974)将四川宝兴标本另立新属--仰鼻猴属(Rhinopithecus,1872=Semnopithecus,1870)以来,其属名一直被分类学家、形态学家和灵长类学家等所承认和采用(Elliot,1913; Hill,1936; Allen,1938;Simpson、1945;Buettner-Janusch,1963;Napier等,1967,等)。     

双管基因枪介导的基因瞬时表达技术在拟南芥中的应用

基因瞬时表达是植物中研究目标基因功能的常用手段。在模式植物拟南芥(Arabidopsis thaliana)中, 相比原生质体和农杆菌介导的基因异源表达技术, 利用粒子轰击进行基因瞬时表达一直鲜有报道。其主要原因是拟南芥叶型相对较小、基因枪操作相对烦琐以及基因表达效率差异较大。该研究通过优化双管基因枪系统, 在营养生长旺盛的拟南芥莲座叶中实现GFP和GUS基因高效表达。同时, 通过GUS报告基因明确了坏死诱导因子BAX、Avh238和ATR13/Rpp13激发拟南芥细胞坏死的表型。但在本氏烟(Nicotiana benthamiana)中明显诱导细胞坏死的Avrblb1/RB基因对, 在拟南芥中却丧失了诱导细胞坏死的活性。由于双管基因枪系统每次轰击时设置平行对照, 可有效降低转化实验中的样本变异度, 为拟南芥及其突变体研究中准确评价基因功能和高通量筛选目标基因提供新的技术参考。     

The Mycobacterium tuberculosis mysB gene product is a functional equivalent of the Escherichia coli sigma factor, KatF

Mycobacterium tuberculosis, the causative agent of tuberculosis, may remain dormant within its host for many years. The nature of this dormant or latent state is not known, but it may be a specialized form of the stationary growth phase. In Escherichia coli, KatF (or RpoS) is the major stationary phase sigma factor regulating an array of genes expressed in this phase of growth. A potential M. tuberculosis katF homologue was cloned using a fragment of the E. coli katF gene as a probe. DNA sequence analysis of a resultant clone showed 100% identity to a fragment of DNA encoding the M. tuberculosis mysA and mysB genes. Overexpression of mysB in M. bovis BCG resulted in an increase in katG mRNA and catalase and peroxidase activity, and an increase in sensitivity of the cells to isoniazid. An increase in katG promoter activity from a reporter vector was demonstrated when mysB was overexpressed from the same plasmid, indicating a direct relationship between MysB and katG expression.     

松嫩平原晚更新世以来古植被演替的初步研究

根据松嫩平原不同地点六个剖面的孢粉分析资料及测年数据, 恢复该区晚更新世晚期以来的六种植被类型, 并且对晚更新世晚期在冰缘气候控制下的三种模式孢粉组合所代表向三种典型植被类型进行讨论。     

The radiation of the Seicercus burkii complex and its congeners (Aves: Sylviidae): molecular genetics and bioacoustics

Cryptic species of passerine birds lack notable morphological differentiation and can best be identified by molecular and bioacoustic markers. Here we investigate seven cryptic species of the golden-spectacled warbler (Seicercus burkii complex) with respect to territorial song and cytochrome-b (cyt-b) sequences. Their phylogenetic relations to other Seicercus species and to members of the genus Phylloscopus are inferred by the same methods. Three separate lineages of Seicercus are nested within different branches of the molecular Phylloscopus tree. The S. burkii complex is a monophyletic unit comprising seven species (S. burkii s. str., S. whistleri, S. valentini, S. soror, S. omeiensis, S. tephrocephalus and S. affinis). S. xanthoschistos turned out to be a close relative of Phylloscopus davisoni within the P. reguloides group. Two isolated sister taxa, S. grammiceps and S. castaniceps, also branch together with the P. reguloides group. Within the S. burkii complex the overall haplotype and nucleotide diversity is highest in taxa from the Chinese middle and upper mountain belt (S. valentini, S. omeiensis and S. soror), indicating at least partially restricted gene flow in these species. This is explained by the fragmentation of high-altitude habitats in China while in the Himalayas the vicariant species S. whistleri inhabits a more continuous mountain belt at the same altitude. For the Chinese species from medium and high altitudes, past range expansion is indicated by significantly negative Tajima Ds. According to pairwise genetic distances, most species of the S. burkii complex have diverged 5 myr ago, the most recent split between S. burkii and S. tephrocephalus is dated 2 myr ago. Coalescence times for haplotype lineages of the different species range from 9 up to 12 myr, and between 5 and 6 myr for S. burkii and S. tephrocephalus.

Within Seicercus divergence of song features such as frequency parameters and syntax structures correlate with genetic distances between taxa. The three cyt-b lineages of Seicercus correspond to different clusters in a discriminant analysis by acoustic parameters. Common syntax structures of territorial song in the Phylloscopus/Seicercus assemblage are: (1) an introductory element derived from specific calls and (2) a syntax of trills and repeated element groups or a combination of both. There are clear indications that these song structures have repeatedly emerged, were lost or were altered in different branches of the phylogenetic tree at different times. Absolute differences between taxa in frequency parameters or in an acoustic divergence index increase significantly with growing genetic distances. However, due to multiple parallel evolution phylogenetic information provided by single acoustic traits decreases with increasing numbers of taxa involved in the investigation.     

Comparison between a native and exotic adelgid as hosts for Laricobius rubidus (Coleoptera: Derodontidae)

Laricobius rubidus LeConte, native to eastern North America whose primary host is Pineus strobi Hartig, has been observed in association with Adelges tsugae, an exotic and lethal pest of hemlock trees in eastern North America. Pineus strobi and A. tsugae were used to evaluate host preference and suitability of this predatory beetle to determine in part its suitability as a potential biological control agent of A. tsugae. In a paired-choice oviposition test, L. rubidus preferred to oviposit in P. strobi ovisacs over those of A. tsugae. However, in the no-choice oviposition test, there was no significant difference in the mean number of eggs laid by L. rubidus females in P. strobi or A. tsugae ovisacs. There were no significant differences in larval developmental time or survivorship for L. rubidus reared on a diet of either P. strobi or A. tsugae. Laricobius rubidus completed development to the adult stage on A. tsugae indicating that it is a suitable host and therefore has the potential to contribute to biological control of A. tsugae in the eastern United States. The introduction of a congener, Laricobius nigrinus to eastern North America may result in competition with L. rubidus on A. tsugae, but the inability of L. nigrinus to complete development on P. strobi suggests that the two congeners will not compete on the primary host of L. rubidus.     

The stability of mutator (MR)-induced X-chromosomal recessive visible mutations in Drosophila melanogaster

In Drosophila, MR (male recombination) second chromosomes are known to act as mutators and recombination inducers in males. The induction of visible mutations by MR is observed at only a limited number of genes, such as singed bristle (sn). raspberry eye colour (ras), yellow body colour (y) and carmine eye colour (car). Futhermore, sn mutations induced by MR are highly unstable, changing from a strong to a weak expression or reverting to the wild-type. It has been hypothesized, by analogy with IS mutations in microorganism, that MR-induced mutations also represent mutations of the insertion type.

In this investigation the stability of two MR-hl22-induced X-linked visible mutations was tested, one singed (sn^MR) and one raspberry (ras^MR). The reversion frequency of both MR-induced mutations was low in the base population as well as upon outcrossing to C(1)DX, y^w f females. The data reported here show that the MR-induced mutations become highly unstable when MR is re-introduced. The change of expression of an MR-induced mutation to a weaker phenotype or to the wild-type occurred at a frequency of 3.3 × 10⁻³ (ras) to 20.4 × 10⁻ (sn).

Recessive lethal mutations induced by MR in the X-chromosomes carrying the MR-inducedsinged or raspberry mutation were isolated and analysed. Among 11 independently MR-induced lethals in the ras^MR-carrying X-chromosome, 4 were found to be allelic to a small deficiency that included the raspberry gene, 13 lethals were induced by MR in the sn^MR-carrying X-chromosome. Of these, 3 were located near the sn locus but none was allelic to a deficiency including the singed gene.     

The bloom-forming ciliate Mesodinium rubrum harbours a single permanent endosymbiont

Whether the red tide Mesodinium rubrum contains a permanent cryptophyte symbiont or whether it only sequesters chloroplasts from cryptophyte prey was addressed using electron microscopy and the dynamics of photosynthesis, chloroplasts and nuclei. Mesodinium rubrum contains a branched cryptophyte symbiont consisting of many chloroplasts, mitochondria, nucleomorphs, an endoplasmic reticulum and one nucleus. The volume of the symbiont constitutes 36% of the consortium and it is separated from its host by a single-cell membrane. The chloroplasts of Mesodium are larger and morphologically different from two Teleaulax species that served as prey. The symbiont nucleus is also much larger than Teleaulax nuclei. Although M. rubrum is functionally a phototroph, sustained growth beyond two to four generations requires ingestion of prey, but less than one prey cell per generation suffices for maximum growth. This suggests that either the ciliate or its symbiont needs an essential growth factor for continuous growth.     

Purification and characterization of the N gene product of bacteriophage lambda

The N protein (pN) specified by bacteriophage λ is an antitermination factor and is required for phage development. pN can be assayed by making use of the observation that the in vitro synthesis of trp mRNA in a reaction programmed with DNA template from λtrp transducing phage bearing N$The N protein (pN) specified by bacteriophage λ is an antitermination factor and is required for phage development. pN can be assayed by making use of the observation that the in vitro synthesis of trp mRNA in a reaction programmed with DNA template from λtrp transducing phage bearing N and fed mutations is pN dependent (Ishii et al., 1980). The assay has been used to purify pN. We have observed that pN forms a complex with E. coll protein(s) and is dissociated in the presence of urea. The complex is not formed in host bacteria bearing thenusA_nusB_ mutations. pN is a basic protein and heat-stable. Using these characteristics, we have purified pN to virtual homogeneity as judged by polyacrylamide gel electrophoresis in the presence of SDS. pN is a monomeric protein and its mol. wt. is approx. 14 000. The antiterminating activity of pNappears to be enhanced by complex formation with host-encoded protein(s) depending on the nusA and/or nusB gene function.      18. Spatial regulation of segment polarity gene expression in the anterior terminal region of the Drosophila blastoderm embryo      Jym Mohler 《Mechanisms of development》1995,50(2-3):151-161 The effects of mutations in five anterior gap genes (hkb, tll, otd, ems and btd) on the spatial expression of the segment polarity genes, wg and hh, were analyzed at the late blastoderm stage and during subsequent development. Both wg and hh are normally expressed at blastoderm stage in two broad domains anterior to the segmental stripes of the trunk region. At the blastoderm stage, each gap gene acts specifically to regulate the expression of either wg or hh in the anterior cephalic region: hkb, otd and btd regulate the anterior blastoderm expression of wg, while tll and ems regulate hh blastoderm expression. Additionally, btd is required for the first segmental stripe (mandibular segment) of both hh and wg at blastoderm stages. The subsequent segmentation of the cephalic segments (preantennal, antennal and intercalary) appears to be dependent on the overlap of the wg and hh cephalic domains as defined by these gap genes at the blastoderm stage. None of these five known gap genes are required for the activation of the labral segment domains of hh and wg, which are presumably either activated directly by maternal pathways or by an unidentified gap gene.      19. 我国主栽食用菌茶薪菇的物种概念      刘祈猛  宋海燕  陈润秀  陈明辉  翟志军  周健平  高扬  胡殿明 《菌物学报》2021,40(5):981-991 茶薪菇是我国广泛栽培的一种食药用菌,由于其形态特征与柱状环伞十分相似,分类地位至今尚不明确.本研究采用形态学和多基因分子系统发育分析相结合的方法,研究茶薪菇和柱状环伞的分类问题,结果表明:茶薪菇和柱状环伞是两个不同但亲缘关系密切的物种;在形态上,茶薪菇菌盖颜色浅褐色至深褐色,油茶味浓,菌环较薄易脱落,褶缘囊状体柱状、烧...      20. 福建漳州水仙花的染色体数目及命名研究   总被引：1，自引：0，他引：1   曾沧江  陈勤娘 《植物研究》1984,4(4):159-164 福建漳州的水仙花分为单瓣水仙及重瓣水仙两类,李时珍早就指出它们乃一物二种[1]。李懋学等(1980)观察了福建漳州、浙江舟山、江苏崇明岛等地的水仙花的染色体数目,均为2n=30,因此认为这三个地区的水仙花都是中国水仙Narcissus tazetta L.var.chinensis Roem.,并从染色体组型分析,阐明它们全是同源三倍体[8]。我们分别观察了漳州的单瓣水仙及重瓣水仙染色体数目,与他们观察的结果有所不同。说明漳州水仙花的染色体数目及其分类问题,有进一步研究之必要。      $

Spatial regulation of segment polarity gene expression in the anterior terminal region of the Drosophila blastoderm embryo

The effects of mutations in five anterior gap genes (hkb, tll, otd, ems and btd) on the spatial expression of the segment polarity genes, wg and hh, were analyzed at the late blastoderm stage and during subsequent development. Both wg and hh are normally expressed at blastoderm stage in two broad domains anterior to the segmental stripes of the trunk region. At the blastoderm stage, each gap gene acts specifically to regulate the expression of either wg or hh in the anterior cephalic region: hkb, otd and btd regulate the anterior blastoderm expression of wg, while tll and ems regulate hh blastoderm expression. Additionally, btd is required for the first segmental stripe (mandibular segment) of both hh and wg at blastoderm stages. The subsequent segmentation of the cephalic segments (preantennal, antennal and intercalary) appears to be dependent on the overlap of the wg and hh cephalic domains as defined by these gap genes at the blastoderm stage. None of these five known gap genes are required for the activation of the labral segment domains of hh and wg, which are presumably either activated directly by maternal pathways or by an unidentified gap gene.     

我国主栽食用菌茶薪菇的物种概念

茶薪菇是我国广泛栽培的一种食药用菌,由于其形态特征与柱状环伞十分相似,分类地位至今尚不明确.本研究采用形态学和多基因分子系统发育分析相结合的方法,研究茶薪菇和柱状环伞的分类问题,结果表明:茶薪菇和柱状环伞是两个不同但亲缘关系密切的物种;在形态上,茶薪菇菌盖颜色浅褐色至深褐色,油茶味浓,菌环较薄易脱落,褶缘囊状体柱状、烧...     

福建漳州水仙花的染色体数目及命名研究

福建漳州的水仙花分为单瓣水仙及重瓣水仙两类,李时珍早就指出它们乃一物二种^[1]。李懋学等(1980)观察了福建漳州、浙江舟山、江苏崇明岛等地的水仙花的染色体数目,均为2n=30,因此认为这三个地区的水仙花都是中国水仙Narcissus tazetta L.var.chinensis Roem.,并从染色体组型分析,阐明它们全是同源三倍体^[8]。我们分别观察了漳州的单瓣水仙及重瓣水仙染色体数目,与他们观察的结果有所不同。说明漳州水仙花的染色体数目及其分类问题,有进一步研究之必要。