香蕉RAPD分析初步研究

比较了不同提取方法对香蕉植株不同部位组织提取 DNA的质量及其 PCR扩增结果 ,对香蕉 RAPD分析中引物种类和浓度 ,复性温度 ,d NTPs,Taq DNA聚合酶浓度 ,热循环数等因素进行了比较影响分析。结果表明 ,虽然改良的 SDS法、CTAB法和 PVP法提取的植株嫩叶和吸芽 DNA提取量和纯度各不相同 ,但其 PCR扩增结果基本相同 ;相同克隆不同植株的 DNA其 PCR扩增结果也基本相同 ;建立了适合香蕉大规模 DNA多态性分析 RAPD反应体系 :2 5 μL反应液中 ,含 1倍缓冲液 ,0 .2 m Md NTPs,0 .3 2 p M随机引物 ,IUTaq酶 ,2 0 ng模板 DNA;反应循环数为 45 ,热循环条件为 94°C,1 min;3 7°C,1 min;72°C,1 .5 min;之前为 94°C,5 min;之后为72°C,1 0 min。在筛选的 2 49个随机引物中 ,有 1 8个在 7个品种上都能扩增出 3～ 1 0条比较清晰条带。     

杜鹃花RAPD条件的优化

以井冈山杜鹃为材料建立了杜鹃花RAPD反应优化体系 ,用于杜鹃花遗传多样性分析 ,以改进的CTAB法提取杜鹃花属植物杜鹃花叶片总DNA ,分别测试了镁离子浓度、dNTP浓度、模板DNA含量、引物浓度、DNA聚合酶量对反应结果的影响。通过各因子的组合比较 ,建立了杜鹃花RAPD优化体系 :2 0 μLPCR反应体积 ,10×Taq酶配套缓冲液 (2 μL) ;1UTaq酶 ;模板DNA10ng ;13.32pmol引物 ;2 .12mmol·L- 1 MgCl2 ;dATP、dCTP、dGTP和dTTP各 0 .15mmol·L- 1 。     

濒危植物七子花RAPD条件的优化

以改进SDS法抽提濒危植物七子花嫩叶总DNA ,进行随机扩增多态DNA(RAPD)分析 ,分别测试了镁离子 ,dNTP ,模板DNA含量 ,引物和DNA聚合酶量对反应结果的影响 ,通过各因子的组合研究 ,可知七子花RAPD分析较适宜的扩增条件是 :1 5μLPCR反应体积 ,1×Taq酶配套缓冲液 (1 0mmol/LTris·HClpH 9.0 ,50mmol/LKCl,0 .1 %TritonX_1 0 0 ) ,2 .5mmol/LMgCl2 ,2UTaq酶 (上海华美公司 ) ,1 0ng模板DNA ,2 0pmol引物 (上海Sangon公司 ) ;dATP、dCTP、dGTP、dTTP各 0 .1mmol/L。     

Hg~(2+)胁迫对浮萍体细胞DNA一级结构和抗氧化酶系的损伤(英文)

主要从DNA一级结构及抗氧化酶系变化两方面 ,研究了Hg2 + 胁迫下浮萍体细胞的损伤。结果表明 :运用随机扩增多态性DNA法 (RandomamplifiedpolymorphismDNA ,RAPD)和DNA梯法 (DNALadder) ,5～ 1 0mg·L- 1 Hg2 +处理组可检测到基因组DNA的明显损伤 ,2 0mg·L- 1 Hg2 + 已导致细胞坏死 ;RAPD法较DNALadder法更灵敏。本文还发现 ,活性氧和抗氧化酶系很可能参与了浮萍体细胞凋亡过程。低浓度的Hg2 + 胁迫可刺激抗氧化酶活性升高 ,以清除体内活性氧 ,而一旦活性氧水平超出一定域值 ,抗氧化酶活性急速下降 ,导致细胞凋亡     

苦丁茶冬青的RAPD影响因素及实验条件的优化

以苦丁茶冬青为材料研究随机扩增多态DNA(RAPD)的影响因素及各种实验条件优化。研究结果表明：模板DNA的浓度适宜范围为20ng／反应-80ng／反应RAPD均可得到一致的结果;dNTVs的适宜浓度范围为200μmol／L-400μmol／L;Mg^2 适宜浓度范围为1．5mmol／L-2．0mmol／L;其合适的复性温度为35—37℃;2min的延伸时间,45次热循环。按照此优化的RAPD条件进行重复实验,实验结果重现性良好,因而确定了苦丁茶冬青RAPD反应体系之最佳的实验条件。     

广霍香的组织培养

1　植物名称　广霍香 (Pogostemoncablin)2　材料类别　嫩叶、嫩枝。3　培养条件　芽诱导与增殖培养基 :( 1 )MS +4PU 30 (四川成都施特优化工厂生产 ) 0 .2 5mg·L- 1 + 3%蔗糖 + 0 .8%琼脂 ;( 2 )MS + 6 BA 0 .5mg·L- 1 + 3%蔗糖 + 0 .8%琼脂。生根培养基 :( 3)MS +NAA 0 .2mg·L- 1 + 1 .5 %蔗糖 + 0 .8%琼脂。上述培养基pH均为5 .8,培养温度为 ( 2 5± 2 )℃ ,光照度为 1 2 0 0lx左右 ,每天光照 1 2h。4　生长与分化情况4.1　不定芽和叶腋丛生芽诱导与增殖　将广霍香嫩枝、嫩叶用自来水冲冼干净后 ,以 0 .1 %升汞表面消毒 1 0m…     

大叶秦艽的组织培养与植株再生

以秦艽的叶和下胚轴为外植体,成功地诱导出愈伤组织和再生植株.诱导愈伤组织最合适的培养基为附加2 m g·L- 1 2 ,4 - D和0 .5 mg·L- 1 6 - BA的MS培养基,诱导率可达到10 0 % .愈伤组织转移到附加2 mg·L- 12 ,4 - D和0 .5 mg·L- 1 KT和5 0 0 m g·L- 1 L H的MS培养基上进行继代培养,增殖后的愈伤组织转移到附加0 .1mg·L- 1 2 ,4 - D和0 .5 m g·L- 1 6 - BA的MS分化培养基上进行分化,其分化率可达到86 .6 7% ,将分化出的芽转接到不加激素的MS上,结果可生长出大量的分化苗.     

线粒体DNA数目不同的人角质形成细胞模型的建立

该研究采用溴化乙锭(ethidium bromide,Et Br)对人永生化角质形成细胞(HaCaT)进行诱导,采用RT-PCR法、MTT法和健那绿染色技术联合鉴定Et Br对HaCaT细胞线粒体DNA(mitochondrial DNA,mt DNA)拷贝数的影响,建立不同mt DNA拷贝数目的HaCaT细胞模型。结果显示,HaCaT细胞经100 ng/m L和50 ng/m L Et Br分别处理10 d后,细胞形态随着处理时间的延长发生改变,由规则铺路石形状逐渐变圆,某些细胞体积变小,成团生长,细胞膜边缘不光滑,且随着mt DNA数目的减少,细胞增殖速率明显下降。RT-PCR分析结果显示,与对照组相比,50 ng/m L和100 ng/m L Et Br处理10 d的HaCaT细胞中,mt DNA的拷贝数分别减少了52.9%和97.6%[以线粒体DNA脱氢酶1(mitochondrial NADH dehydrogenase 1 gene,ND1)与甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)的比值来表示mt DNA的相对拷贝数变化情况]。健那绿染色显示,对照组HaCaT细胞中可见分散存在的蓝绿色线粒体颗粒,100 ng/m L Et Br处理10 d后的HaCaT细胞中,显微镜下蓝绿色线粒体颗粒明显减少。综上所述,该研究证明了HaCaT细胞可通过Et Br诱导被成功培养成ρ–细胞,且Et Br浓度与HaCaT细胞mt DNA拷贝数的变化在一定的范围内存在着剂量–效应关系。     

Hg2+胁迫对浮萍体细胞DNA一级结构和抗氧化酶系的损伤(英文)

 主要从DNA一级结构及抗氧化酶系变化两方面,研究了Hg2+胁迫下浮萍体细胞的损伤。结果表明：运用随机扩增多态性DNA法（Random amplified polymorphism DNA, RAPD）和DNA梯法（DNA Ladder）,5～10 mg·L-1　Hg2+处理组可检测到基因组DNA的明显损伤,20 mg·-1 Hg2+已导致细胞坏死;RAPD法较DNA Ladder法更灵敏。本文还发现,活性氧和抗氧化酶系很可能参与了浮萍体细胞凋亡过程。低浓度的Hg2+胁迫可刺激抗氧化酶活性升高,以清除体内活性氧,而一旦活性氧水平超出一定域值,抗氧化酶活性急速下降,导致细胞凋亡。      

珍珠菜提取物抗肿瘤作用的初步研究

观察灌服珍珠菜提取物对小鼠肉瘤 ( S1 80 )的抑制作用 ,用相同方法跟踪珍珠菜的抗肿瘤有效部位。用紫外分光光度法 ,观察了珍珠菜提取物对环磷酰胺造成骨髓抑制的保护作用 ;采用 MTT法观察不同浓度珍珠菜提取物对人白血病 HL - 6 0及 K 5 6 2细胞的体外抑制作用。结果表明珍珠菜浸膏 ( 40 0 0 m g/ kg· d)、石油醚部位 ( 2 0 0 m g/kg· d)、氯仿部位 ( 2 0 0 m g/ kg· d)、乙酸乙酯 ( 2 0 0 mg/ kg· d)部位、正丁醇部位 ( 2 0 0 mg/ kg· d)及水溶性部位 ( 2 0 0mg/ kg· d)对小鼠肉瘤 ( S1 80 )抑瘤率分别为 5 2 .74%,16 .5 1%,2 8.41%,42 .17%,48.2 1%,2 5 .2 4%;小鼠灌服环磷酰胺 ( 2 5 mg/ kg· d)能引起骨髓抑制而使骨髓中 DNA含量下降 ,如同时灌服珍珠菜浸膏 ( 2 0 0 0 m g/ kg· d)则可使小鼠骨髓 DNA含量恢复至接近正常水平 ;珍珠菜总黄酮提取物 ( 2 10μg/ m L )、乙酸乙酯部位 ( 180μg/ m L )、正丁醇部位 ( 2 10μg/ m L )作用 48h对 HL - 6 0细胞的抑制率分别为 35 .6 6 %,45 .85 %,42 .36 %;IC5 0分别为 92 .85 ,16 4.93,40 2 .6 1μg/ m L ;对 K5 6 2细胞抑制率为 5 9.94%,70 .34%,77.98%;IC5 0分别为 17.6 4,5 2 .5 3,6 4.70μg/ m L。因此 ,珍珠菜提取物对小鼠肉瘤 ( S180 )有显     

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.