Study on fibroin heavy chain of the silkwormBombyx mori by fluorescencein situ hybridization (FISH)

The sericulture industry plays a very important role in our national economy. Silkworm (Bombyx mori) is always regarded as a model animal and biological reactor. There have been detailed studies on the structure, expression and control and molecular evolution of silk genes. However, few, if any, reports are available on the localization of structural genes in silkworm by molecular cytogenetics. The present experiment has tentatively localized theFib-H gene at the distal end of the 25th linkage group, namely at the 25-0.0 position, and verified thatFib-H has only one locus, thus providing a temporary solution to the problem about its localization.     

Study on fibroin heavy chain of the silkwormBombyx mori by fluorescencein situ hybridization (FISH)

The sericulture industry plays a very important role in our national economy. Silkworm (Bombyx mori) is always regarded as a model animal and biological reactor. There have been detailed studies on the structure, expression and control and molecular evolution of silk genes. However, few, if any, reports are available on the localization of structural genes in silkworm by molecular cytogenetics. The present experiment has tentatively localized theFib-H gene at the distal end of the 25th linkage group, namely at the 25-0.0 position, and verified thatFib-H has only one locus, thus providing a temporary solution to the problem about its localization.     

树木抗病基因研究进展

在长期进化过程中,植物形成了一系列防御机制,抵抗各类病原物入侵,抗病R基因在其中发挥着关键作用。R基因的研究,在农作物中取得很大进展,已成为植物病理学的研究热点。相比之下,树木抗病基因研究较为落后,虽从苹果、杨树和柚子中已分离了几个与已知R基因具有类似结构与功能的基因,但还没有真正的树木抗病R基因被克隆出来;目前,大部分研究主要集中在利用分子标记构建连锁图谱,寻找抗病数量性状位点（Quantitative trait loci,QTL）和与R基因紧密连锁或共分离的质量性状标记;其中一些标记已经应用在分子辅助选育中,并显示了诱人的应用前景。另外,利用已知抗病R基因的保守区域,从多种植物中已扩增出许多抗病基因类似序列,它们大多被转化为与R基因紧密连锁的标记或被当作抗病候选基因。     

Intercellular communication and carcinogenesis

Two types of intercellular communication (humoral and cell contact-mediated) are involved in control of cellular function in multicellular organisms, both of them mediated by membrane-embedded proteins. Involvement of aberrant humoral communication in carcinogenesis has been well documented and genes coding for some growth factors and their receptors have been classified as oncogenes. More recently, cell contact-mediated communication has been found to have an important role in carcinogenesis, and some genes coding for proteins involved in this type of communication appear to form a family of tumor-suppressor genes. Both homologous (among normal or (pre-)cancerous cells) as well as heterologous (between normal and (pre)cancerous cells) communications appear to play important roles in cell growth control. Gap junctional intercellular communication (GJIC) is the only means by which multicellular organisms can exchange low molecular weight signals directly from within one cell to the interior of neighboring cells. GJIC is altered by many tumor-promoting agents and in many human and rodent tumors. We have recently shown that liver tumor-promoting agents inhibit GJIC in the rat liver in vivo. Molecular mechanisms which could lead to aberrant GJIC include: (1) mutation of connexin genes; (2) reduced and/or aberrant expression of connexin mRNA; (3) aberrant localization of connexin proteins, i.e., intracytoplasmic rather than in the cytoplasmic membrane; and (4) modulation of connexin functions by other proteins, such as those involved in extracellular matrix and cell adhesion. Whilst mutations of the cx 32 gene appear to be rare in tumors, cx 37 gene mutations have been reported in a mouse lung tumor cell line. Our results suggest that aberrant connexin localization is rather common in cancer cells and that possible molecular mechanisms include aberrant phosphorylation of connexin proteins and lack of cell adhesion molecules. Studies on transfection of connexin genes into tumor cells suggest that certain connexin genes (e.g., cx 26, cx 43 and cx 32) act as tumor-suppressor genes.     

普通烟草ARF基因家族序列的鉴定与表达分析

ARF(AUXIN RESPONSE FACTOR)基因含有一个B3功能域和具有转录激活或抑制活性的中心功能域,在植物发育过程中起到非常重要的作用。本研究采用生物信息学方法,根据拟南芥ARF基因序列鉴定了普通烟草基因组中的ARF基因,并对家族成员进行了序列特征、系统发生、亚细胞定位和表达模式分析。目前在普通烟草基因组中共得到50个ARF基因成员,其基因结构相对复杂,一般含有10个外显子。亚细胞定位结果表明,少数ARF蛋白定位到线粒体或叶绿体,大多数未检测到定位信号。转录组数据分析表明,ARF基因具有不同的组织表达模式,部分基因表现出组织特异性。这些研究结果为普通烟草ARF基因家族功能的深入研究奠定了基础。     

小麦高分子量谷蛋白亚基及其基因的研究进展

主要介绍了小麦高分子量谷蛋白亚基（HMW－GS）及其基因的研究进展情况,目前,转基因小麦的技术已经逐渐成熟,由于分子生物学领域分子标记技术的迅速发展,尤其是PCR技术的广泛应用,为实现外源优良储藏蛋白基因导入改良品种提供了可能,利用已知小麦品种的基因序列设计引物,从众多的未知小麦品种中扩增出新基因加以研究并做外源优质HMW－GS基因的转入已成为一种趋势。     

An Improved Nonfluorescent Detection System for in Situ Hybridization in Plants

Molecular cytogenetics, particularly the localization of DNA sequences by in situ hybridization, has increased our understanding about the genomic structure of plants and animals. We demonstrate here the application of an improved nonfluorescent in situ hybridization system detection (DAKO GenPoint system) to plant chromosomes. Using this system, highly repetitive 18S-25S rRNA genes were mapped on Vicia faba chromosomes (2n = 12). The modified method of this horseradish peroxidase based enzymatic detection system gave satisfactory results that are comparable to fluorescent signal detection.     

Classical and molecular cytogenetics of the zebrafish, Danio rerio (Cyprinidae, Cypriniformes): an overview

The zebrafish, Danio rerio, has recently become the model system for the genetic analysis of vertebrate development. This paper reviews the advances in zebrafish cytogenetics, obtained through classical and molecular techniques, which will lead to the assignment of specific linkage groups to specific chromosome pairs in the zebrafish genome project. Several chromosome pairs of the 50-chromosome karyotype of D. rerio were differentially stained by classical staining techniques and additional information has been obtained by molecular cytogenetics. Indeed, the analysis of constitutive heterochromatin by C-banding and base-specific fluorochrome staining had suggested a differential composition of peri- and paracentromeric constitutive heterochromatin. The chromosome mapping of distinct AT- and GC-rich zebrafish satellite DNAs by means of PRINS (Primed in situ) and multicolor FISH (Fluorescence in situ Hybridization) has confirmed this hypothesis, which therefore provided the chromosome localization of 10% of the zebrafish genome. The analysis of nucleolus organizer regions (NORs) by silver staining and by FISH with 18S rDNA has also revealed the existence of variable and inactive NORs, in addition to those on the terminal regions of the long arms of the three NOR-bearing chromosome pairs. Other multicopy genes, such as minor ribosomal genes, or multicopy repeats, such as telomere specific sequences, have now been mapped on zebrafish chromosomes. The latest advancement in zebrafish molecular cytogenetics is the chromosome mapping of single locus genes. Single-copy genes from each of the 25 genetic linkage groups are now being mapped on zebrafish chromosomes by using PAC clones.     

互花米草NAC转录因子家族的鉴定与表达分析

互花米草(Spartina alterniflora)作为一种海岸带盐生植物,高度耐盐胁迫,但因为缺少参考基因组,其耐盐的分子机制却尚未见报道。NAC家族蛋白是植物特有的转录因子,调控植物的生长发育和胁迫应答。为了鉴定互花米草NAC蛋白(SaNAC)并探究它们与互花米草生长发育及胁迫响应之间的关系,本研究以互花米草三代全长转录组数据为参考,通过与水稻(Oryza sativa)、拟南芥(Arabidopsis thaliana)和玉米(Zea mays)NAC蛋白序列进行比对,并结合保守功能域进一步筛选,最终找到62个SaNAC蛋白。从蛋白序列比对、进化、motif预测、同源性比较、亚细胞定位、组织表达以及非生物胁迫下的基因差异表达等方面分别对互花米草NAC家族成员进行分析,结果发现SaNAC蛋白均含有保守的NAM结构域,且在进化上与水稻NAC家族具有一定的相似性;SaNAC家族中的两个蛋白SaNAC9和SaNAC49在细胞核表达;另外,本研究还发现互花米草SaNAC基因表达具有高度组织和胁迫应答差异性。这些结果表明互花米草NAC转录因子家族不仅具有保守的功能域,而且在调控互花米草的生长发育和非生物胁迫响应过程中具有重要的作用。     

Gene transfer and gene mapping in mammalian cells in culture

Summary The ability to transfer mammalian genes parasexually has opened new possibilities for gene mapping and fine structure mapping and offers great potential for contributing to several aspects of mammalian biology, including gene expression and genetic engineering. The DNA transferred has ranged from whole genomes to single genes and smaller segments of DNA. The transfer of whole genomes by cell fusion forms cell hybrids, which has promoted the extensive mapping of human and mouse genes. Transfer, by cell fusion, of rearranged chromosomes has contributed significantly to determining close linkage and the assignment of genes to specific chromosomal regions. Transfer of single chromosomes has been achieved utilizing microcells fused to recipient cells. Metaphase chromosomes have been isolated and used to transfer single-to-multigenic DNA segments. DNA-mediated gene transfer, simulating bacterial transformation, has achieved transfer of single-copy genes. By utilizing DNA cleaved with restriction endonucleases, gene transfer is being employed as a bioassay for the purification of genes. Gene mapping and the fate of transferred genes can be examined now at the molecular level using sequence-specific probes. Recently, single genes have been clones into eucaryotic and procaryotic vectors for transfer into mammalian cells. Moreover, recombinant libraries in which entire mammalian genomes are represented collectively are a rich new source of transferable genes. Methodology for transferring mammalian genetic information and applications for mapping mammalian genes is presented and prospects for the future discussed. Presented in the symposium on Gene Transfer, Differentiation and Neoplasia in Plant and Animal Cells at the 30th Annual Meeting of the Tissue Culture Association, Seattle, Washington, June 10–14, 1979. This symposium was supported in part by Grant CA 26748 from the National Cancer Institute, DHEW, and Grant RD-67 from the American Cancer Society. Supported by NIH grants HD 05196 and GM 20454 and by MOD grants 1-485 and 1-692.     

抑制素基因的研究进展

抑制素是性腺分泌的一种糖蛋白激素,它具有抑制垂体促卵泡素合成和分泌的作用。本文介绍了抑制素α亚基基因（INHA）、抑制素βA亚基基因（INHBA）、抑制素βB亚基基因（INHBB）的克隆、结构、定位、多态性、表达、分子调节及其与繁殖性能和癌症的关系。绵羊INHA、INHBA和INHBB基因分别被定位到2q41→q43、4q26和2q31→q33。INHA、INHBA和INHBB基因对绵羊产羔数都有显著的影响。抑制素βB亚基基因突变的雌性小鼠有明显的发育和繁殖缺陷。INHA 基因与妇女卵巢早衰显著相关。 Abstract：Inhibins are gonadal glycoprotein hormones belonging to the transforming growth factor-βsuperfamily that act to suppress pituitary follicle-stimulating hormone synthesis and secretion. In this paper, we briefly introduced the cloning, structure, localization, polymorphism, expression, molecular regulation of inhibin-α(INHA), -βA (INHBA) and -βB (INHBB) subunit genes and their relationships with reproductive performance and cancer. The inhibin genes (INHA, INHBA and INHBB) had significant effect on litter size in sheep. The ovine INHA, INHBA and INHBB genes had been mapped to chromosomes 2q41→q43, 4q26 and 2q31→q33, respectively. The female mice carrying INHBB mutations suffered from distinct developmental and reproductive defects. The INHA gene was significantly associated with premature ovarian failure in women.     

水稻逆境相关转录因子研究进展

干旱、盐碱、高温和低温等逆境因子胁迫水稻的生长发育,进而影响水稻的产量和品质。因此,研究水稻的抗逆性,尤其是揭示其抗逆分子机理具有重要的生物学意义。近年来,水稻抗逆分子机理的研究主要集中在转录因子及其分子调控机制方面。在水稻中,目前研究较多的转录因子类型主要有b ZIP、MYB/MYC、WRKY、AP2/EREBP和NAC,它们的结构通常由DNA结合结构域、转录活化结构域、寡聚化位点和核定位信号组成。转录因子在水稻逆境信号转导途径中起着中心调节作用,它们将逆境信号传递和放大,通过与目的基因启动子区中顺式作用元件特异结合,调控下游多个逆境相关基因的表达,从而引起水稻对逆境应答反应,最终实现水稻获得综合抗逆性的提升。该文简要概述了植物转录因子的调控机制、结构特点、分类与功能特性,重点论述了转录因子在水稻抗逆中的作用,指出了转录因子应用过程中转基因水稻产生的负效应问题,并提出了解决负效应问题的研究思路,同时展望了今后转录因子的研究前景,以期为挖掘和应用新的水稻转录因子基因以及阐明其抗逆调控机制提供理论依据。     

基于COⅠ和Cyt b基因序列的凤蝶科六属分子系统学研究

对凤蝶科6属25种的COⅠ基因和20种Cyt b基因的部分序列进行测定和分析, 探讨它们之间的系统发育关系; 以茶小卷叶蛾Adoxophyes honmai为外群, 用邻接法(neighbor-joining, NJ)、 最大简约法(maximum parsimony, MP)和贝叶斯法(Bayesian inference, BI)重建了凤蝶科6属的分子系统树。结果表明： COⅠ基因部分序列长度为661 bp, 其中保守位点417个, 可变位点244个, 简约信息位点191个; A+T的平均含量为70.3%, 明显高于C+G的平均含量29.6%。Cyt b基因部分序列长度为433 bp, 其中保守位点239个, 可变位点194个, 简约信息位点135个; A+T的平均含量为74.2%, 明显高于C+G的平均含量25.7%。分子系统树表明, 凤蝶属Papilio、 斑凤蝶属Chilasa、 尾凤蝶属Bhutanitis、 珠凤蝶属Pachliopta和喙凤蝶属Teinopalpus为单系性, 与传统形态分类结果相一致。但青凤蝶属Graphium单系性不够明确, 需要进一步探讨。研究结果为我国凤蝶科分子系统学研究积累了资料。     

Toward the complete genomic map and molecular pathology of human chromosome 4

The identification of disease genes via molecular DNA cloning has revolutionized human genetics and medicine. Both the candidate gene approach and positional cloning have been used successfully. The defects causing Huntington's disease, facioscapulohumeral muscular dystrophy, piebaldism, Hurler/Scheie syndrome, one form of autosomal recessive retinitis pigmentosa, and a second locus for autosomal dominant polycystic kidney disease have recently been localized to chromosome 4. In addition to the rapid progress in the cloning of the 203-megabase chromosome, the presence of more than 60 closely spaced microsatellites on this chromosome will undoubtedly lead to the localization of additional disease genes. In order to consider cloned genes as potential candidates for disorders assigned to chromosome 4, it is important to collect and order all genes with respect to their chromosomal localization. Analysis of cytogenetically visible interstitial and terminal deletions should also be helpful in defining new disease gene loci and in mapping novel genes. These data represent the status quo of the integrated molecular map for chromosome 4.     

葡萄Actin基因家族的鉴定及进化和表达分析

采用生物信息学方法从葡萄(Vitis vinifera Linn.)全基因组中鉴定Actin基因家族,并对各基因的染色体定位和结构特征,编码蛋白质的理化性质、亚细胞定位、二级结构、三级结构和系统进化,以及不同组织的基因表达进行研究.结果表明:葡萄Actin基因家族16个基因分布在12条染色体上.16个基因的结构特征及其编码蛋白质的理化性质差异较大.16个基因的长度及其内含子总长度的变化范围较大,编码序列(CDS)和外显子总长度的变化范围较小.除登录号GSVIVG01008254001和GSVIVG01014035001的基因外,其他14个基因的GC含量均低于其CDS的GC含量.除登录号GSVIVG01008254001的基因外,其他15个基因编码的蛋白质的理论相对分子质量为12534.54~82612.33,理论等电点为pI 4.92~pI 9.13.16个基因编码蛋白质的消光系数为14105~73645,脂肪族氨基酸指数为65.54~92.06,其中9个为稳定蛋白,7个为不稳定蛋白.除登录号GSVIVG01014035001的基因外,其他15个基因编码的蛋白质均为亲水性蛋白.登录号GSVIVG01016517001的基因编码的蛋白质定位于细胞质和细胞核,其他15个基因编码的蛋白质定位于细胞质.二级结构和三级结构显示:葡萄Actin基因家族16个基因编码的蛋白质均由α螺旋、无规则卷曲和延伸链构成,且总体以无规则卷曲为主.系统进化分析和不同组织的基因表达分析结果显示:与拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕相似,葡萄Actin基因家族16个基因编码的蛋白质分为3个亚家族,ClassⅡ亚家族(营养型)包括登录号GSVIVG01003099001和GSVIVG01026580001的基因编码的蛋白质,这2个基因在所有组织中的表达均较高;ClassⅢ亚家族(生殖型)包括登录号GSVIVG01033494001、GSVIVG01024980001和GSVIVG01016550001的基因编码的蛋白质,这3个基因在花粉、雄蕊和花中的表达均较高;ClassⅠ亚家族包括其他11个基因编码的蛋白质,这11个基因在各组织中的表达总体上较低.研究结果显示:葡萄Actin基因家族的表达具有组织特异性.     

An apparent interlocus gene conversion-like event at a putative tumor suppressor gene locus on human chromosome 6q27 in a Burkitt's lymphoma cell line.

A region of minimal deletion in B-cell non-Hodgkin's lymphoma (B-NHL) has recently been defined between D6S186 and D6S227 spanning 5-9 Mb at 6q26-q27, predicting the presence of at least one tumor suppressor gene (TSG) at this locus. During the construction of a deletion map in the B-NHL tumor panel, we report the identification of a Burkitt's lymphoma cell line, BL74, having an apparent homozygous deletion at the D6S347 locus, internal to the critical region. Since this case may facilitate the localization of the target TSG, a detailed structural molecular characterization and search for candidate genes were undertaken at this locus. While BL74 underwent a loss of heterozygosity at 6q26-q27, D6S347 was also likely subjected to a somatic interlocus gene conversion-like event between two homologous but distinct loci, resulting in the homozygous replacement of a 1860- to 2067-bp segment of one locus with the corresponding segment copied from the other locus. Two genes at this locus were identified, but their lack of expression in B-cell lineages tentatively excludes them as candidate TSGs. Another still unidentified gene at this locus may be disrupted by the gene conversion-like event, which would represent a novel mechanism of TSG inactivation.