丙型肝炎病毒RNA打点杂交检测方法同RT－PCR方法的比较

采用ＨＣＶ基因组结构区Ｃ区ｃＤＮＡ探针和非结构区ＮＳ３－４区ｃＤＮＡ探针,建立了用打点杂交（ｄｏｔｂｌｏｔｈｙｂｒｉｄｉｚａｔｉｏｎ）检测血清中ＨＣＶＲＮＡ的方法,同采用ＨＣＶ基因组５’端非编码区的一对寡核苷酸引物通过逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）检测血清中ＨＣＶＲＮＡ的方法相比较,发现两种方法都能快速早期和特异地检出血清中ＨＣＶＲＮＡ,但ＲＴ－ＰＣＲ法敏感性优于ＲＮＡ打点杂交法。对于无血清学指标的慢性ＮＡＮＢ肝炎病人的诊断,可采用这两种方法。这两种方法的敏感性在很大程度上依赖于引物和探针的敏感性,以及ＲＮＡ提取方法。ＲＴ－ＰＣＲ法适用于诊断病毒血症和复制,打点杂交法适用于研究ＨＣＶＲＮＡ量的变化,对治疗的评价,以及为实验筛选较高滴度的ＨＣＶＲＮＡ阳性样本。     

啤酒酵母DNA修复基因RAD24温度敏感突变株的分离及其特性的初步研究

以ＹＣｐｌａｃ系列带Ｔｒｐ、Ｈｉｓ和Ｕｒａ标志基因的载体为骨架构建含野生型和经羟胺处理的突变型的啤酒酵母ＲＡＤ２４基因质粒,用质粒替换方法分离ＲＡＤ２４基因温度敏感突变株（ｒａｄ２４－ｔｓ３）．紫外生存试验发现,ｒａｄ２４－ｔｓ３对紫外线敏感;同位素（３Ｈ－ＴｄＲ,３Ｈ－ＵＲ,３Ｈ－Ｌｅｕ）参入试验表明,该突变株ＤＮＡ、ＲＮＡ及蛋白质合成均较野生型明显降低．     

α1肾上腺素受体亚型在家兔胸主动脉平滑肌细胞生长中的作用

本实验在体外培养的家兔胸主动脉平滑肌细胞上,利用＾３Ｈ－Ｔｈｙｍｉｄｉｎｅ（＾３Ｈ－ＴｄＲ）掺入的方法,观察了α１－肾上腺素受体（ａｄｒｅｎｅｒｇｉｃ ｒｅｃｅｐｔｏｒ,ＡＲ）及其亚型对平滑肌细胞ＤＮＡ合成的影响。结果显示：去甲肾上腺素激动α１－ＡＲ能促进平滑肌细胞ＤＮＡ合成,并有以下特点：（１）α１Ａ激动促进平滑肌细胞ＤＮＡ合成,但α１Ｂ激动抑制这一合成作用;（２）α１Ａ促进ＤＮＡ合成的作用与ｃ     

粉防己碱对血管紧张素Ⅱ促进心肌成纤维细胞增殖的抑制作用

血管紧张素（ＡＮＧ）Ⅱ对新生大鼠无血清培养第二代心肌成纤维细胞（ＦＢ）具有剂量依赖性的促进增殖、ＤＮＡ和ＲＮＡ以及胶原合成的作用。ＡＮＧⅡ１０７ｍｏｌ／Ｌ浓度时,ＦＢ的［３Ｈ］胸腺嘧啶核苷（Ｔｈｙｍｉｄｉｎｅ）,［３Ｈ］尿嘧啶核苷（Ｕｒｉｄｉｎｅ）,［３Ｈ］脯氨酸（Ｐｒｏｌｉｎｅ）掺入较对照组分别增加１６７％,５８％,１７４％,此刺激作用可被特异性ＡＮＧⅡ受体拮抗剂阻断;粉防己碱预处理也可抑制ＡＮＧⅡ上述作用,且呈剂量依赖性。以上结果提示：ＡＮＧⅡ通过受体介导直接刺激心肌ＦＢ增殖和胶原合成,引起心肌纤维化;钙拮抗剂粉防己碱对ＡＮＧⅡ导致的心肌纤维化具有抑制作用。     

雀麦花叶病毒缺陷型RNA_（3b）的研究

采用ＰＥＧ沉淀和差速离心的方法提纯雀麦花叶病毒的Ｇ和Ｔ分离物。利用蛋白酶Ｋ和两相酚法制备雀麦花叶病毒的总ＲＮＡｓ。将Ｇ和Ｔ分离物ＲＮＡｓ进行琼脂糖凝胶电泳,结果发现Ｂｒ－ＭＶ－Ｇ除含有正常的ＲＮＡ组分外,还出现了另一新的ＲＮＡ＿（３ｂ）组分,其分子量为０．５０×１０￣６。ＲＮＡ＿（３ｂ）只出现在大麦寄主中,而在昆诺基上缺失。ＲＮＡ＿（３ｂ）仅依靠于其来源的Ｇ分离物的ＲＮＡｓ进行复制。以ＲＮＡ＿（３ｂ）为模板合成￣（３２）Ｐ－ｃＤＮＡ探针,和ＢｒＭＶ－Ｇ分离物的ＲＮＡｓ进行分子杂交试验表明：ＲＮＡ＿（３ｂ）属ＲＮＡ＿３的缺陷型组分,它依赖于ＢｒＭＶ－Ｇ－ＲＮＡ＿３的帮助才能在大麦寄主中复制。ＲＮＡ＿（３ｂ）的出现和缺失对ＢｒＭＶ的症状表现没有影响。     

乙型肝炎病人重叠感染丙型肝炎的评价

应用ＥＬＩＳＡ和ＰＣＲ法检测５０２例乙肝病人血清,４０１例ＨＢｓＡｇ阳性血清中,有１１４例（２８．４％）抗－ＨＣＶ和ＨＣＶＲＮＡ双项阳性,２５例（６．２％）ＨＣＶＲＮＡ单项阳性;２１例（５．２％）抗－ＨＣＶ单项阳性。将ＨＢｓＡｇ乙肝病人分成ＨＢＶＤＮＡ,ＨＢｅＡｇ阳性组和ＨＢＶＤＮＡ,ＨＢｅＡｇ阴性组。前者抗－ＨＣＶ阳性率为１１．６％～２０．５％,ＨＣＶＲＮＡ阳性率为１６．２％～２０．５％。后者抗－ＨＣＶ阳性率为２０．２％～５５．６％,ＨＣＶＲＮＡ阳性率为２３％～６０．３％。结果说明长期携带ＨＢＶ者和慢性乙肝病人均可重叠ＨＣＶ感染。ＨＢＶＤＮＡ阳性组抗－ＨＣＶ和ＨＣＶＲＮＡ阳性率明显高于ＨＢＶＤＮＡ阳性组     

对硫磷对三角褐指藻核酸和蛋白质合成动态的影响

应用同位素标记法研究了对磷磷对三角褐指灌核酸和蛋白质合成动态的影响。结果表明,低浓度的对硫酸（≤１．５ｍｇ／Ｌ）对三角褐指的生长有刺激作用,而高浓度的对硫磷（≥２．０ｍｇ／Ｌ）却严重抑制三角褐指藻的生长,低浓度划硫磷在促进生长的过程中,藻细胞中蛋白质、ＤＮＡ、ＲＮＡ３种大分子物质的合成活跃,其合成速度升高,而在高浓度对硫磷的胁迫下,蛋白质,ＤＮＡ,ＲＮＡ的合成明显地受到了抑制,合成速度降低。     

有序差异显示:一种基因表达谱系统比较法

系统研究具有同一基因组的各种细胞群之间基因的差异表达谱十分重要。目前,研究基因差异表达的技术大致有ｍＲＮＡ差异显示［１］、ＲＤＡ［３］、ＳＳＨ［４、５］和ｃＤＮＡ阵列［６］等。近几年,还发展了一些研究基因差异表达谱系统的技术,如ＲＬＣＳ（ｒｅｓｔｒｉｃｔｉｏｎｌａｎｄ－ｍａｒｋｃＤＮＡｓｃａｎｎｉｎｇ）［８］、ＧＥＦ（ｇｅｎｅｅｘｐｒｅｓ－ｓｉｏｎｆｉｎｇｅｒｐｒｉｎｔｉｎｇ）［２］和ＲＮＡ指纹法［９］等。然而,这些技术或较为复杂,或灵敏度偏低。本文拟介绍一种有效的基因表达谱系统比较法——有序差…     

神经节苷脂GM_3对人白血病J6－2细胞磷脂酰胆碱代谢的影响

神经节苷脂ＧＭ３诱导人单核样白血病Ｊ６－２细胞沿单核／巨噬细胞途径分化．在ＧＭ３诱导分化同时,Ｊ６－２细胞磷脂代谢发生了显著变化．采用（（３２）Ｐ）Ｐｉ、［ＧＨ３－３Ｈ］胆碱和［ＣＨ３－３Ｈ］ＳＡＭ参入实验对ＧＭ３影响Ｊ６－２细胞ＰＣ代谢的机制进行了初步的探讨．ＧＭ３促进［（３２）Ｐ］Ｐｉ参入Ｊ６－２细胞ＰＣ;抑制［ＣＨ３－３Ｈ］胆碱参入ＰＣ及ＰＣ合成的前体磷酸胆碱及ＣＤＰ－胆碱;ＧＭ３促进［ＣＨ３－３Ｈ］ＳＡＭ参入ＰＣ,但抑制［ＣＨ３－３Ｈ］ＳＡＭ参入ＰＣ合成的前体胆碱、磷酸胆碱和ＣＤＰ－胆碱．上述结果提示,ＧＭ３抑制Ｊ６－２细胞ＰＣ合成的ＣＤＰ－胆碱途径,促进ＰＣ合成的ＰＥ甲基化途径．     

花生种子内源ABA含量变化及其与活力的关系

花生（Ａｒａｃｈｉｓ ｈｙｐｏｇａｅａ Ｌ．）种子发育过程中,胚轴内源ＡＢＡ 含量一直是增加的;种皮内源ＡＢＡ含量在果针入土后４０ ｄ 最大,然后急剧下降;子叶内源ＡＢＡ 含量在果针入土后６０ ｄ 出现高峰,然后有轻微下降。种子活力指数和萌发时内源ＡＢＡ 的净下降量有密切关系。甘露醇可促进离体胚内源ＡＢＡ 合成,１－甲基－３－苯基－５（３－［三氟甲基］－苯基－４－（１氢）－吡啶）抑制子叶内源ＡＢＡ 的合成,子叶和胚轴存在不同的ＡＢＡ 合成途径。种子早熟和早萌处理时,内源ＡＢＡ 含量都下降,胚轴在种子由发育向萌发转换中起着十分重要的作用     

表油菜素内酯对绿豆上胚轴内源IAA及其氧化酶的影响

用0.5ppm表油菜素内酯处理绿豆幼苗,显著促进上胚轴伸长生长,若切除真叶则可抑制表油菜素内酯诱导的效应。三碘苯甲酸(TIBA)也可抑制表油菜素内酯促进的伸长生长。外源IAA能部分恢复TIBA的抑制效应。经处理的上胚轴内源IAA含量明显高于对照。暗示表油菜素内酯可能通过对内源IAA的调节来促进绿豆上胚轴的伸长生长。 表油菜素内酯处理的绿豆上胚轴组织中,与生长素降解密切相关的IAA氧化酶以及过氧化物酶活性均明显低于对照。     

A proteomics study of the mung bean epicotyl regulated by brassinosteroids under conditions of chilling stress

Mung bean CYP90A2 is a putative brassinosteroid (BR) synthetic gene that shares 77% identity with the Arabidopsis CPD gene. It was strongly suppressed by chilling stress. This implies that exogenous treatment with BR could allow the plant to recover from the inhibited growth caused by chilling. In this study, we used proteomics to investigate whether the mung bean epicotyl can be regulated by brassinosteroids under conditions of chilling stress. Mung bean epicotyls whose growth was initially suppressed by chilling partly recovered their ability to elongate after treatment with 24-epibrassinolde; 17 proteins down-regulated by this chilling were re-up-regulated. These up-regulated proteins are involved in methionine assimilation, ATP synthesis, cell wall construction and the stress response. This is consistent with the re-up-regulation of methionine synthase and S-adenosyl-L-methionine synthetase, since chilling-inhibited mung bean epicotyl elongation could be partially recovered by exogenous treatment with DL-methionine. This is the first proteome established for the mung bean species. The regulatory relationship between brassinosteroids and chilling conditions was investigated, and possible mechanisms are discussed herein.     

切除上胚轴对绿豆子叶衰老逆转及其核酸和蛋白质代谢的效应

在绿豆子叶衰老达到不归点(萌发后5～6d)前切除上胚轴可使开始衰老的子叶中核酸和蛋白质含量回升,衰老短期逆转。衰老不归点后的子叶中核酸和蛋白质变化的主要特征是:丧失了较多的核主带DNA、25S、18S rRNA以及大部分可溶性蛋白质组分,一种小分子DNA 组分完全消失。不归点前切除上胚轴可使上述核酸和蛋白质组分明显增加,表明子叶衰老的逆转可能与这些重要功能物质的回升有关。在切除上胚轴的茎顶涂抹IAA,能阻止子叶核酸和蛋白质回升,也消除了切除上胚轴对子叶裹老的逆转作用。     

绿豆子叶衰老期间核酸代谢的变化

萌发绿豆的子叶自然衰老期间,核酸含量降低,RNA降低的幅度比DNA大。电泳分析结果表明,子叶衰老期间细胞核主带DNA明显降低;而迁移慢的卫星带DNA变化不大。在RNA各组分中,18S rRNA从衰老前期就开始降低;25S rRNA和4～5S小分子RNA到衰老后期才缓慢下降。DNase和RNase活性在子叶整个衰老期间都明显升高,是导致核酸含量下降的主要原因。~3H-核苷掺入试验表明,核酸的合成速率在子叶衰老前期有所上升,到衰老后期又降低。poly(A)~ -mRNA含量在子叶开始衰老时明显上升。     

Modulation of Chlorophyll, Carotene and Xanthophyll Formation by Penicillin, Benzyladenine and Embryonic Axis in Mung Bean (Phaseolus aureus L.) Cotyledons

Penicillin stimulated the synthesis of pigments in the cotyledonsof intact embryos and excised cotyledons of mung bean (Phaseolusaureus L.) and enhanced benzyladenine-induced accumulation ofchloroplast pigments. The presence of the embryonic axis duringlight exposure proved to be beneficial for chlorophyll synthesisby the cotyledons whereas its presence in dark germination producedan adverse effect. The possible involvement of nucleic acidand protein synthesis in light-regulated chlorophyll formationis suggested. The stimulating effect on pigment synthesis providedby penicillin in this system seems to involve a maintenanceof nucleic acid and protein synthesis. Phaseolus aureus L., mung bean, pigment synthesis, cotyledons     

Comparison of the effects of epibrassinolide and steroidal estrogens on adventitious root growth and early shoot development in mung bean cuttings

Concentrations of 24-epibrassinolide as low as 0.1 μ M consistently inhibited adventitious root formation and elongation in both hypocotyl and epicotyl cuttings from mung bean ( Phaseolus aureus L.). Similar, but less pronounced, inhibitory effects on root elongation were also observed with estrone sulphate and estradiol sulphate. With regards to root number, estrone sulphate enhanced this in both types of cutting, whereas estradiol sulphate was stimulatory in hypocotyl cuttings but inhibitory in epicotyl cuttings. Brassinolide caused a marked stimulation of epicotyl (but not hypocotyl) elongation and a swelling and splitting of the epicotyl in both types of cutting, whereas estrogens varied in their effect from inhibition of epicotyl growth to no effect. Root-applied brassinolide and estrogen sulphates brought about similar morphological abnormalities in shoots viz. epinasty and inrolling of primary leaves and delayed expansion of the first trifoliate leaf.     

DNA synthesis in the elongating nondividing cells of the lentil epicotyl and its promotion by gibberellin

Two-day-old lentil seedlings, (Lens culinaris Med.) were incubated for a 48-hour period with and without gibberellin (GA) in the presence and absence of 5-fluorodeoxyuridine (FUDR). The number of cells per epicotyl did not increase during this period. Growth of the epicotyl was thus due to cell elongation alone.

The elongating cells of this tissue synthesized DNA. GA promoted and FUDR inhibited cell elongation, DNA synthesis, and RNA synthesis in the tissue.

FUDR promoted uptake of thymidine and thymidine incorporation into cellular DNA, presumably by inhibiting synthesis of endogenous thymidine. Presence of GA promoted thymidine incorporation into cellular DNA and uridine incorporation into cellular RNA. In either case, there was no effect on the uptake of the precursor into the tissue.

Fractionation of thymidine-labeled nucleic acids on a MAK column showed that thymidine was exclusively incorporated into the DNA fraction. Presence of GA promoted thymidine incorporation into this fraction and also increased the amount of ribosomal RNA.

The data provide direct evidence for the conclusion that DNA synthesis is necessary for elongation of certain plant cells.

     

Involvement of cellulose synthesis in actions of gibberellin and kinetin on cell expansion. 2,6-Dicholorobenzonitrile as a new cellulose-synthesis inhibitor

In azuki bean epicotyl sections, 2,6-dichlorobenzonitrile inhibitedcellulose synthesis but showed no effect on the synthesis ofnon-cellulose wall materials and trichloroacetic-acid-insolublecytoplasmic substances. Dichlorobenzonitrile reversed gibberellinpromotion and kinetin inhibition of auxin-induced epicotyl elongationwithout affecting auxin-induced elongation, itself. (Received August 17, 1973; )