The Ah Phenotype. Survey of Forty-Eight Rat Strains and Twenty Inbred Mouse Strains

Forty-four inbred and four randombred rat strains and 20 inbred mouse strains were examined for their Ah phenotype by determining the induction of liver microsomal aryl hydrocarbon (benzo[a]pyrene) hydroxylase activity (EC 1.14.14.1) by intraperitoneal treatment with either β-naphthoflavone or 3-methylcholanthrene. All 48 rat strains were found to be Ah-responsive. The maximally induced hydroxylase specific activities of the ALB/Pit, MNR/Pit, MR/Pit, SHR/Pit, and Sprague-Dawley strains were of the same order of magnitude as the basal hydroxylase specific activities of the ACI/Pit, F344/Pit, OKA/Pit, and MNR/N strains. Six of the 20 mouse strains were Ah-nonresponsive (i.e. lacking the normal induction response and presumably lacking detectable amounts of the Ah receptor). The basal hydroxylase specific activities of the BDL/N, NFS/N, STAR/N, and ST/JN mouse strains were more than twice as high as the maximally induced hydroxylase specific activity of the CBA/HT strain.——To date, 24 Ah-nonresponsive mouse strains have been identified, out of a total of 68 known to have been characterized. The reasons for not finding a single Ah-nonresponsive inbred rat strain—as compared with about one Ah-nonresponsive inbred mouse strain found for every three examined—remain unknown.     

Comparative Sequence Analysis of the Non-Protein-Coding Mitochondrial DNA of Inbred Rat Strains

The proper function of mammalian mitochondria necessitates a coordinated expression of both nuclear and mitochondrial genes, most likely due to the co-evolution of nuclear and mitochondrial genomes. The non-protein coding regions of mitochondrial DNA (mtDNA) including the D-loop, tRNA and rRNA genes form a major component of this regulated expression unit. Here we present comparative analyses of the non-protein-coding regions from 27 Rattus norvegicus mtDNA sequences. There were two variable positions in 12S rRNA, 20 in 16S rRNA, eight within the tRNA genes and 13 in the D-loop. Only one of the three neutrality tests used demonstrated statistically significant evidence for selection in 16S rRNA and tRNA-Cys. Based on our analyses of conserved sequences, we propose that some of the variable nucleotide positions identified in 16S rRNA and tRNA-Cys, and the D-loop might be important for mitochondrial function and its regulation.     

转SAMS基因玉米自交系获得及抗旱性分析

以玉米自交系‘掖478’的茎尖为受体,采用农杆菌介导法将小麦抗旱SAMS基因转入玉米中,用PCR和RT-PCR法对转化玉米进行检测,并以18%PEG-6000模拟水分胁迫对T1代转基因玉米和非转基因玉米进行抗旱性分析。结果显示:(1)共转化518个玉米芽,得到453株转化苗。获得T0代阳性植株16株,转化率为3.53%,有14株自交结实,经RT-PCR检测,T1代转基因玉米有9个株系为稳定遗传阳性株系。(2)在同一水分胁迫时间下,转基因玉米的叶片相对含水量、叶绿素含量、脯氨酸含量和SOD活性均高于非转基因玉米,而转基因玉米叶片的电导率、MDA含量均低于非转基因玉米。在60 h水分胁迫处理下,转基因玉米叶片相对含水量、叶绿素含量、脯氨酸含量和SOD活性比非转基因玉米上升8.16%、20.02%、32.21%、22.77%,而转基因玉米叶片的电导率、MDA含量比非转基因玉米下降14.38%、29.41%。研究表明,通过导入SAMS基因,可以提高玉米的抗旱性。     

玉米优异早熟种质单330开花相关性状的QTL分析

玉米开花相关性状与玉米的成熟期和产量有密切的联系。通过对玉米CN165×单330(早熟种质)群体的130个F2:3家系开花相关性状在3个环境下进行分子鉴定和数量性状位点(QTL)分析,结果表明,在3个环境中检测到控制抽雄天数的10个QTL,分别位于第2、3、4、5、7、8染色体上,在第8染色体上同一区域在3种环境下都检测到了QTL;检测到控制散粉天数的10个QTL,分别位于第1、2、3、5、7、8染色体上,在第8染色体上同一区域在2种环境下都检测到了QTL;检测到控制吐丝天数的4个QTL,分别位于第4、5、8染色体上,在第8染色体不同环境下都检测到了2个QTL;仅仅在一个环境中检测到控制ASI的2个QTL,分别位于第6、9染色体上。这些QTL的基因效应以部分显性和超显性为主。研究表明,第8染色体上ph i060-um c2401区域(8.03~8.04)是一个研究开花相关性状的重要基因组区段,涉及到的标记可以作为分子标记辅助选择的重要候选标记。     

乙脑病毒持续感染株preM区序列分析

为了研究乙型脑炎病毒持续感染株preM区域基因序列变异及其意义,我们将两种乙脑病毒野生株(JaGAr-01株和Nakayama株)分别感染人肝癌KN73细胞,经过多次细胞传代后建立乙脑病毒持续感染模型,收集感染细胞经反复冻融获取变异病毒.利用preM区特异引物进行RT-PCR法得到两种病毒的preM区基因片段,应用基因测序反应进行序列分析,并对两种病毒株preM区序列进行比较.preM区基因测序结果显示,与JaGAr-01野生株比较,JaGAr-01持续感染变异株(JaG-per)有1个核苷酸上碱基发生变异(第26位U→G)并导致相应氨基酸发生置换(第9位亮氨酸→精氨酸);Nakayama持续感染变异株(Nak-per)与其野生株相比则有11个核苷酸上碱基存在差异(第26位U→G,第37位G→A,第39位C→U,第45位U→C,第51位U→C,第99位U→C,第126位U→C,第165位C→U,第189位C→U,第195位C→U,第198位U→C),但仅有其中第26位、第37位、第39位的碱基变异引起相应编码的氨基酸发生置换(第9位亮氨酸→精氨酸及第13位缬氨酸→异亮氨酸).对比还发现变异后的JaGAr-01持续感染株与Nakayama持续感染株的基因序列相同.认为乙脑病毒持续感染变异株preM区存在基因变异,这种变异可能与该区参与病毒持续感染及维持病毒生物学特性有关.     

乙脑病毒持续感染株preM区序列分析

为了研究乙型脑炎病毒持续感染株preM区域基因序列变异及其意义,我们将两种乙脑病毒野生株(JaGAr-01株和Nakayama株)分别感染人肝癌KN73细胞,经过多次细胞传代后建立乙脑病毒持续感染模型,收集感染细胞经反复冻融获取变异病毒。利用preM区特异引物进行RT-PCR法得到两种病毒的preM区基因片段,应用基因测序反应进行序列分析,并对两种病毒株preM区序列进行比较。preM区基因测序结果显示,与JaGAr-01野生株比较,JaGAr-01持续感染变异株(JaG-per)有1个核苷酸上碱基发生变异(第26位U→G)并导致相应氨基酸发生置换(第9位亮氨酸→精氨酸);Nakayama持续感染变异株(Nak-per)与其野生株相比则有11个核苷酸上碱基存在差异(第26位U→G,第37位G→A,第39位C→U,第45位U→C,第51位U→C,第99位U→C,第126位U→C,第165位C→U,第189位C→U,第195位C→U,第198位U→C),但仅有其中第26位、第37位、第39位的碱基变异引起相应编码的氨基酸发生置换(第9位亮氨酸→精氨酸及第13位缬氨酸→异亮氨酸)。对比还发现变异后的JaGAr-01持续感染株与Nakayama持续感染株的基因序列相同。认为乙脑病毒持续感染变异株preM区存在基因变异,这种变异可能与该区参与病毒持续感染及维持病毒生物学特性有关。     

The Physical and Genetic Framework of the Maize B73 Genome

Maize is a major cereal crop and an important model system for basic biological research. Knowledge gained from maize research can also be used to genetically improve its grass relatives such as sorghum, wheat, and rice. The primary objective of the Maize Genome Sequencing Consortium (MGSC) was to generate a reference genome sequence that was integrated with both the physical and genetic maps. Using a previously published integrated genetic and physical map, combined with in-coming maize genomic sequence, new sequence-based genetic markers, and an optical map, we dynamically picked a minimum tiling path (MTP) of 16,910 bacterial artificial chromosome (BAC) and fosmid clones that were used by the MGSC to sequence the maize genome. The final MTP resulted in a significantly improved physical map that reduced the number of contigs from 721 to 435, incorporated a total of 8,315 mapped markers, and ordered and oriented the majority of FPC contigs. The new integrated physical and genetic map covered 2,120 Mb (93%) of the 2,300-Mb genome, of which 405 contigs were anchored to the genetic map, totaling 2,103.4 Mb (99.2% of the 2,120 Mb physical map). More importantly, 336 contigs, comprising 94.0% of the physical map (∼1,993 Mb), were ordered and oriented. Finally we used all available physical, sequence, genetic, and optical data to generate a golden path (AGP) of chromosome-based pseudomolecules, herein referred to as the B73 Reference Genome Sequence version 1 (B73 RefGen_v1).     

Mating-Type Mutations in SCHIZOSACCHAROMYCES POMBE: Isolation of Mutants and Analysis of Strains with an h or h Phenotype

Mutants defective in various steps of the sexual cycle have been isolated from homothallic strains of Schizosaccharomyces pombe by Bresch, Müller and Egel (1968). These mutants include heterothallic h(+) and h(-) strains. We have isolated additional h(+) and h(- ) mutants from homothallic strains. Those mutants which are due to mutations in the mating-type region were analyzed in detail. Our results show that the mating-type gene mat2 not only has a function in copulation and meiosis, but that it also regulates the formation of the map1 gene product (map1 is a mating-type auxiliary gene). Some of the h( -) mutants have lost only one of the three functions while others are defective in at least two, and perhaps all three, functions. Further, we show that the mat1(-) allele of h(90) strains can mutate to mat1(+) but that mutations in mat2 appear to affect the mutational behavior of mat1. Finally, we describe a new inactive mating-type allele, mat2*, which is different from mat2(0) in that it can mutate to mat2(+).     

Human Immunodeficiency Virus Neurotropism: an Analysis of Viral Replication and Cytopathicity for Divergent Strains in Monocytes and Microglia

Productive replication of human immunodeficiency virus type 1 (HIV-1) in brain macrophages and microglia is a critical component of viral neuropathogenesis. However, how virus-macrophage interactions lead to neurological disease remains incompletely understood. Possibly, a differential ability of virus to replicate in brain tissue macrophages versus macrophages in other tissues underlies HIV-1 neurovirulence. To these ends, we established systems for the isolation and propagation of pure populations of human microglia and then analyzed the viral life cycles of divergent HIV-1 strains in these cells and in cultured monocytes by using identical viral inocula and indicator systems. The HIV-1 isolates included those isolated from blood, lung tissue, cerebrospinal fluids (CSF), and brain tissues of infected subjects: HIV-1_ADA and HIV-1_89.6 (from peripheral blood mononuclear cells), HIV-1_DJV and HIV-1_JR-FL (from brain tissue), HIV-1_SF162 (from CSF), and HIV-1_BAL (from lung tissue). The synthesis of viral nucleic acids and viral mRNA, cytopathicity, and release of progeny virions were assessed. A significant heterogeneity among macrophage-tropic isolates for infection of monocytes and microglia was demonstrated. Importantly, a complete analysis of the viral life cycle revealed no preferential differences in the abilities of the HIV-1 strains tested to replicate in microglia and/or monocytes. Macrophage tropism likely dictates the abilities of HIV-1 to invade, replicate, and incite disease within its microglial target cells.     

玉米P25自交系抗锈病基因的遗传分析及SSR分子标记定位

以玉米南方型锈病免疫自交系P2 5和感病自交系F3 4 9及F1、F2 、B1和B2 为材料 ,采用主基因 多基因混合遗传模型研究了P2 5的抗病遗传规律。结果表明 :自交系P2 5的抗病基因为一主基因 ,表现为加性效应 ,没有检测出多基因 ,其在F2 、B1和B2 群体的遗传率分别为 81 88%、38 14 %和 5 5 1%。利用SSR分子标记技术 ,以组合P2 5×F3 4 9的F2 :3 家系作为构图群体 ,构建了玉米SSR遗传连锁图谱 ,并将玉米抗南方型锈病基因定位于 10号染色体上 ,与phi0 5 9标记的遗传距离为 5 8cM。     

Phenotype and Genetics of Progressive Sensorineural Hearing Loss (Snhl1) in the LXS Set of Recombinant Inbred Strains of Mice

Progressive sensorineural hearing loss is the most common form of acquired hearing impairment in the human population. It is also highly prevalent in inbred strains of mice, providing an experimental avenue to systematically map genetic risk factors and to dissect the molecular pathways that orchestrate hearing in peripheral sensory hair cells. Therefore, we ascertained hearing function in the inbred long sleep (ILS) and inbred short sleep (ISS) strains. Using auditory-evoked brain stem response (ABR) and distortion product otoacoustic emission (DPOAE) measurements, we found that ISS mice developed a high-frequency hearing loss at twelve weeks of age that progressed to lower frequencies by 26 weeks of age in the presence of normal endocochlear potentials and unremarkable inner ear histology. ILS mice exhibited milder hearing loss, showing elevated thresholds and reduced DPOAEs at the higher frequencies by 26 weeks of age. To map the genetic variants that underlie this hearing loss we computed ABR thresholds of 63 recombinant inbred stains derived from the ISS and ILS founder strains. A single locus was linked to markers associated with ISS alleles on chromosome 10 with a highly significant logarithm of odds (LOD) score of 15.8. The 2-LOD confidence interval spans ∼4 Megabases located at position 54–60 Mb. This locus, termed sensorineural hearing loss 1 (Snhl1), accounts for approximately 82% of the phenotypic variation. In summary, this study identifies a novel hearing loss locus on chromosome 10 and attests to the prevalence and genetic heterogeneity of progressive hearing loss in common mouse strains.     

中国骨干玉米自交系抗旱性分析与评价

在新疆典型干旱、半干旱地区,通过玉米自交系的14个主要产量性状对干旱胁迫的响应特征进行分析,用主成分分析法筛选其抗旱性的主要鉴定指标,并采用抗旱系数和抗旱指数对自交系的抗旱性进行聚类分析,综合评价47份自交系的抗旱性。结果显示:(1)干旱导致玉米自交系株高、穗位高、叶绿素含量降低,果穗短小,秃尖增大,穗行数和行粒数减小,千粒重和穗粒重降低,最终造成产量下降。(2)各自交系抗旱性强弱与干旱条件下的穗粒重、穗粗、行粒数、千粒重、株高、穗位高、穗长、秃尖长、穗行数、叶绿素含量等性状具有较密切的关系,其中抗旱性表现较好的自交系是掖52106、英64、478等,其次是Mo17Ht、Mo17、丹340、郑58、X178、C103等。研究表明,在育种实践中,田间自交系抗旱性筛选应以穗粒重、穗粗、行粒数、千粒重、株高、穗位高、穗长、秃尖长、穗行数、叶绿素含量等为主要指标,并结合产量抗旱系数、抗旱指数对自交系抗旱性进行综合评价。     

玉米FAD2基因的克隆及序列分析

高等植物中的A12脂肪酸脱饱和酶是将油酸转化为亚油酸的酶。根据已发表的其他高等植物的FAD2基因的保守序列设计同源引物,通过RT—PCR从玉米幼胚中扩增得到一个特异的cDNA基因片段。通过生物信息学分析,从玉米幼胚cDNA和基因组中均扩增得到1164 bp FAD2基因（GenBank登陆号：DQ496227）,它编码387个氨基酸,含有完整的ORF框,在ORF框内无内含子。序列联配与树状分析结果表明,FAD2推导的氨基酸序列与其他物种的A12脱饱和酶基因具有同源性。它含有3个组氨酸保守域和2段很长的疏水区,是一个跨膜4次的膜结合蛋白。半定量RT—PCR分析显示FAD2基因在玉米幼胚中表达量最高,在叶、茎、根中亦有低水平表达。     

Proteomic Analysis of Silk Viability in Maize Inbred Lines and Their Corresponding Hybrids

A long period of silk viability is critical for a good seed setting rate in maize (Zea mays L.), especially for inbred lines and hybrids with a long interval between anthesis and silking. To explore the molecular mechanism of silk viability and its heterosis, three inbred lines with different silk viability characteristics (Xun928, Lx9801, and Zong3) and their two hybrids (Xun928×Zong3 and Lx9801×Zong3) were analyzed at different developmental stages by a proteomic method. The differentially accumulated proteins were identified by mass spectrometry and classified into metabolism, protein biosynthesis and folding, signal transduction and hormone homeostasis, stress and defense responses, and cellular processes. Proteins involved in nutrient (methionine) and energy (ATP) supply, which support the pollen tube growth in the silk, were important for silk viability and its heterosis. The additive and dominant effects at a single locus, as well as complex epistatic interactions at two or more loci in metabolic pathways, were the primary contributors for mid-parent heterosis of silk viability. Additionally, the proteins involved in the metabolism of anthocyanins, which indirectly negatively regulate local hormone accumulation, were also important for the mid-parent heterosis of silk viability. These results also might imply the developmental dependence of heterosis, because many of the differentially accumulated proteins made distinct contributions to the heterosis of silk viability at specific developmental stages.     

Comparison of Three Inbred Mouse Strains with Respect to General and Genetic Radiosensitivity

The radiosensitivities of three mouse strains (BALB/cLacY, C3H/SnY, and 101/HY) have been compared using the following parameters: survival after irradiation at a dose of 6–7 Gy, chromosome aberration frequency in bone marrow cells after irradiation at a dose of 1.5 Gy, and the change in testis weight and frequency of abnormal sperm heads (ASHs) after irradiation at doses from 0.5 to 4 Gy. Strain BALB/c is the most radiosensitive with respect to the survival and chromosome aberration frequency in the bone marrow but the most resistant with respect to the change in testis weight and the frequency of abnormal sperm heads. Strain 101/HY was the most resistant with respect to survival and chromosome aberration frequency in bone marrow after irradiation but the most radiosensitive with respect to testis damage.     

Genome Analysis of Listeria monocytogenes Sequence Type 8 Strains Persisting in Salmon and Poultry Processing Environments and Comparison with Related Strains

Listeria monocytogenes is an important foodborne pathogen responsible for the disease listeriosis, and can be found throughout the environment, in many foods and in food processing facilities. The main cause of listeriosis is consumption of food contaminated from sources in food processing environments. Persistence in food processing facilities has previously been shown for the L. monocytogenes sequence type (ST) 8 subtype. In the current study, five ST8 strains were subjected to whole-genome sequencing and compared with five additionally available ST8 genomes, allowing comparison of strains from salmon, poultry and cheese industry, in addition to a human clinical isolate. Genome-wide analysis of single-nucleotide polymorphisms (SNPs) confirmed that almost identical strains were detected in a Danish salmon processing plant in 1996 and in a Norwegian salmon processing plant in 2001 and 2011. Furthermore, we show that L. monocytogenes ST8 was likely to have been transferred between two poultry processing plants as a result of relocation of processing equipment. The SNP data were used to infer the phylogeny of the ST8 strains, separating them into two main genetic groups. Within each group, the plasmid and prophage content was almost entirely conserved, but between groups, these sequences showed strong divergence. The accessory genome of the ST8 strains harbored genetic elements which could be involved in rendering the ST8 strains resilient to incoming mobile genetic elements. These included two restriction-modification loci, one of which was predicted to show phase variable recognition sequence specificity through site-specific domain shuffling. Analysis indicated that the ST8 strains harbor all important known L. monocytogenes virulence factors, and ST8 strains are commonly identified as the causative agents of invasive listeriosis. Therefore, the persistence of this L. monocytogenes subtype in food processing facilities poses a significant concern for food safety.     

猪圆环病毒2型新疆株全基因组的滚环扩增与序列分析

滚环扩增技术是一种体外恒温DNA扩增方法,特异性好,敏感性强,已广泛应用于微生物学领域。首先采用鉴别PCR对来自新疆某地区的5份猪病料进行猪圆环病毒2型的检测,接着对检出的2份猪圆环病毒2型阳性DNA样品进行滚环扩增。滚环扩增产物经单一限制性内切酶(SacⅡ)酶切及琼脂糖凝胶电泳鉴定,结果显示2份样品都出现了猪圆环病毒2型基因组大小的条带。对目的条带进行回收、克隆与测序,结果表明2株新疆株猪圆环病毒2型的全基因组大小皆为1768bp。遗传进化分析显示2株的基因型为PCV2a和PCV2e。     

Association of the Recombination-Deficient Phenotype of Bacillus subtilis recC Strains with the Presence of an SPO2 Prophage

The recombination-defective phenotype associated with the recC genetic locus in Bacillus subtilis is not due to a chromosomal mutation at this site but rather to the presence of an integrated SPO2 prophage.     

玉米S组细胞质雄性不育线粒体R区序列与多型性分析

玉米Ｓ组细胞难性不育（ＣＭＳ）可能与线粒体基因组中的Ｒ区域有关。对不同核背景下唐徐、双２种Ｓ胞质的线粒体ＤＮＡ以Ｒ区特异探针的Ｓｏｕｔｈｅｒｎ分析发现均有６．７ｋｂ、４．５ｋｂ、１．８ｋｂ的３条谱带,分别对应于２种位于线粒体基因组中间的类型和１个线性末端,并且核背景对这３种不同形式的Ｒ区域的量有影响。对Ｍｏ１７和７７核背景下Ｎ、Ｔ、Ｃ４种胞质１７种材料的玉米线粒体基因组中Ｒ区的Ｓｏｕｔｈｅｒｎ分析