完整植物细胞的电击基因转移

本文研究了高压电脉冲对大白菜悬浮细胞通透性和成活率的影响。与原生质体比较,完整细胞可以耐受更高幅值的电脉冲。采用45℃热激,酸性介质(pH4.0)或巯基乙醇预处理黄瓜、水稻和小麦完整植物细胞,有效地提高了导入率。于黄瓜和水稻未去壁的完整细胞,成功地实现了外源 cAT(氯霉素乙酰转移酶)基因的电击导入和瞬间表达。     

脉冲电泳不同参数对介导GFP基因转化玉米种胚效果的研究

利用脉冲电泳介导绿色荧光蛋白(GFP)基因导入玉米种胚;以GFP基因在种胚中瞬时表达作为外源基因导人种胚细胞的标记,分析了外源．DNA浓度、电泳时间、电压、电流转换时间等脉冲电泳转化参数对种胚发芽率和外源基因导入率的影响。结果表明：脉冲电泳时间对种胚发芽率和外源GFP基因导入率影响最大;通过脉冲电泳可将外源基因导入胚芽细胞,其GFP基因导人种子的频率与各电泳参数均呈二次曲线关系,300μg／ml外源DNA浓度、120min电泳时间、5V电压、2s电流转换时间可作为脉冲电泳介导玉米种胚转化较适宜的参数。     

CAT和GUS基因在水稻玉米等作物原生质体中的瞬间表达

使用电击法和 PEG 法将 CAT 和 GUS 基因分别与 CaMV 35S 启动子构成的两种嵌合基因,转移到水稻、玉米、烟草、胡萝卜和香石竹(Dianthus caryophyllus)等品种的原生质体中,均得到了瞬间表达。在电击法中,大多数转化后玉米、烟草和胡萝卜原生质体表现出明显的基因活性。转化效率与所用仪器的电击参数有关。PEG 转化中 PEG 处理时间较国外报道稍有缩短,但转化后的水稻、烟草、胡萝卜和香石竹样品中均得到了阳性结果。     

基因枪在水稻遗传转化中的应用及其转化技术的优化

1983年Zambryski等人用根瘤农杆菌介导法进行烟草基因转移,获得了世界上首例转基因植株.随后,应用DNA直接导入技术如电击法(electroporation)和PEG介导法(PEG—mediated)成功地获得了转基因水稻植株.近年来,随着基因枪技术的建立和发展,水稻遗传转化成功的报道逐年增多.目前基因枪技术在植物遗传转化中的应用超过了根瘤农杆菌介导和其它转化方法的应用.这是因为基因枪转化技术不受植物种类的限制,不需要以原生质体作为转化的受体,可以将外源基因直接导入细胞、组织或器官,因而克服了根瘤农杆菌     

外源GUS基因在PEG介导的花椰菜原生质体中的瞬间表达

为了将外源基因导入花椰菜原生质体获得转基因植株,本文研究了ＰＥＧ介导的外源基因在花椰菜下胚轴原生质体中的瞬间表达。（１）２０％ＰＥＧ将质粒ｐＢＩ２２１导入原生质体后ＧＵＳ表达比１３％ＰＥＧ导入的高,但易造成原生质体损伤。（２）热激处理增强表达,但在随后的培养过程中易造成原生质体降解。（３）原生质体状况对表达有重要影响,５ｄ龄下胚轴原生质体比８ｄ龄的表达强。（４）不同质粒及启动子表达强度不同。质粒ｐＫＩＷＩ１０１比ｐＢＩ２２１表达强３倍左右。     

外源GUS基因在PEG介导的花椰菜原生质体中的瞬间表达

为了将外源基因导入花椰菜原生质体获昨转基因植株,本文研究了ＰＥＧ介导的外源基因在花椰菜一上胚轴原生质体中的瞬间表达。（１）２０％ＰＥＧ将质粒ｐＢＩ２２１导入原生质体后ＧＵＳ表达比１３％ＰＥＧ导入的高,但易造成原生体损伤。（２）热激处理增强表达,但在随后的培养过程中易造成原生质体降解。（３）原生质体状况对表达有重要影响,５ｄ龄下胚轴原生质体比８ｄ龄的表达强。（４）不同质粒及启动子表达强度不同,质粒ｐ     

外源亚精胺对盐胁迫下黄瓜幼苗体内抗氧化酶活性的影响

以不同耐盐性黄瓜品种“长春密刺”和“津春2号”为材料,采用营养液栽培,研究了外源亚精胺(Spd)对NaCl胁迫下黄瓜幼苗叶片与根系中超氧阴离子(O2-.)产生速率、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性的影响。结果表明,外源Spd对未经盐胁迫处理(对照)黄瓜幼苗体内O2-.产生速率、SOD、CAT和POD活性均无显著性影响;盐胁迫处理提高了O2-.产生速率,SOD、POD和CAT活性都有不同程度的升高;外源Spd处理进一步提高了盐胁迫下SOD、POD和CAT活性,减缓了O2-.产生速率。与耐盐型“长春密刺”品种相比,盐胁迫对盐敏感型“津春2号”影响较大,外源Spd对盐敏感型黄瓜品种盐胁迫伤害的缓解作用较大。表明盐胁迫下外源Spd可缓解盐胁迫对膜的伤害,从而提高黄瓜幼苗的耐盐性。     

玉米原生质体经工程操作转移基因后成株

据报道,培育重组体作物又取得一个重大突破。美国山道士作物保护公司(加州帕洛阿尔托)的研究人员成功地把一个外源基因转移入玉米原生质体,并将此原生质体培育成完整植株。他们是用电击穿孔法把一抗卡那霉素基因插入玉米原生质体,转化效率为典型性的0.5～1％。关键步骤是将经转化的原生质体培养在一层“饲喂者”细胞上,使之长成愈伤组织。这些饲喂者细胞是一种液体悬浮液中的黑色墨西哥甜玉米(Black Mexican Swe-     

激光微束照射引起高等植物细胞的遗传变异

除少数植物的原生质体转化较容易外,高等植物的外源基因导入极为困难。特别是许多单子叶作物的原生质体培养和再生很困难,因此人们期望把克隆基因直接导入细胞来避免大量的组织培养,激光微束可穿透植物细胞壁因而能促进细胞对基因的吸收。     

影响枯草杆菌原生质体转化的因素

原生质体转化是将外源基因导入细菌的主要方法之一,其转化频率受制于多种因素。本研究以激BacillussubtilisDB104为宿主菌,以枯草杆菌的高表达型质粒pNQ122为外源DNA,研究了原生质体再生率、原生质体浓度和用于制备原生质体的细胞生长期对转化频率的影响,获得了在该系统中实现高频率转化的条件。该转化条件使外源基因在多个B.subtilis菌株中的转化成为可能,并使从B.subtilis中筛选中性蛋白酶基因获得成功。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.