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61.
几种濒危植物及其近缘类群总DNA的提取与鉴定 总被引:122,自引:0,他引:122
用低pH 介质,高盐沉淀蛋白质方法成功地从银杉(Cathaya argyrophylla Chun etKuang)、矮牡丹(Paeonia suffruticosa var. spontanea)、南川升麻(Cim icifuga nanchuanensisHsiao)、裂叶沙参(Adenophora lobophylla)的同属种泡沙参(A. potaninii)等植物中提取和部分纯化了细胞总DNA,并对其产率、质量和纯度作了鉴定。此方法的关键是用了一个低pH提取介质,它能有效防止组织破碎及沉淀大量材料时的电离化作用及酚化合物的进一步氧化。所得DNA 不需经氯化铯梯度离心或柱层析,直接可用于限制性片断长度多态性(RFLP)及随机扩增的DNA多态性(RAPD)等分子水平的遗传标记。为检测濒危植物的遗传多样性提供了一套迅速、简便和可靠的技术方案 相似文献
62.
Shukuro Araki Shigehiro Yi Tatsufumi Murakami Susumu Watanabe Shinichi Ikegawa Kiyoshi Takahashi Ken-ichi Yamarnura 《Molecular neurobiology》1994,8(1):15-23
To analyze the pathologic processes of amyloid deposition in type I familial amyloidotic polyneuropathy (FAP), mice were made
transgenic by introducing the human mutant transthyretin (TTR) gene(MT-hMet 30). An inbred strain of mouse, C57 BL/6, was
chosen. Transgenic mice were killed using ether anesthesia at 3-mo intervals up to 24 mo after birth. In these transgenic
mice, amyloid deposition started in the gastrointestinal tract, cardiovascular system, and kidneys and extended to various
other organs and tissues with advancing age. The pattern of amyloid deposition was similar to that observed in human autopsy
cases of FAP, except for its absence in the choroid plexus and in the peripheral and autonomic nervous systems.
We extracted the amyloid fibrils from kidneys of these mice with a human mutant TTR gene and analyzed them immunochemically
and electronmicroscopically. Deposited amyloid was shown to be composed of human mutant TTR and mouse serum amyloid P component.
Amyloid fibril from transgenic mice was morphologically and immunohistochemically similar to that of human FAP.
The most striking pathologic feature of the transgenic mice was the absence of amyloid deposition in the peripheral and autonomic
nervous tissues. Thus, other intrinsic factors may be involved in amyloid deposition in the nervous tissues of human FAP. 相似文献
63.
大血藤科植物的分类学研究 总被引:4,自引:0,他引:4
作者对大血藤科植物的花性、叶片内部结构形态、花粉形态、染色体核型及过氧化物酶和脂酶同工酶进行了比较研究,结果表明:1)大血藤科植物外形上的两性花,其雄蕊的形态退化、花药始终不开裂,为功能上的雌花,因此其花为单性,同株,同序或异序;12)首次指出大血藤属植物的染色体数为2n=2x=22,属小型染色体,与木通科植物2n=2x=32、30、28的染色体数明显不同,支持Stapf(1926)将其从木通科分出另立为科的观点;3)单叶和复叶可出现于同一植株上,而且在叶片内部结构、花粉形态、染色体核型、过氧化物酶及脂酶同工酶等性状上,大血藤与单叶血滕间均无实质性的差异,因此将单叶血藤归并于大血藤中。 相似文献
64.
Yi Sun Mahendra S. Rao Richard E. Zigmond Story C. Landis 《Developmental neurobiology》1994,25(4):415-430
Vasoactive intestinal peptide (VIP) expression increases in sympathetic neurons when they are grown in dissociated cell or explant cultures and when they are axotomized in vivo. In dissociated cell culture, the magnitude of the VIP increase was reduced when nonneuronal cells were removed and medium conditioned by ganglionic nonneuronal cells increased VIP in neuron-enriched cultures. Antiserum Against cholinergic differentiation factor (also leukemia inhibitory factor; CDF/LIF), but not against ciliary neurotrophic factor, immunoprecipitated this activity. Medium conditioned by sympathetic ganglion explants also contained a VIP-stimulatory molecule that was immunoprecipitated by CDF/LIF antiserum, and CDF/LIF antiserum partially blocked VIP induction in explants. CDF/LIF mRNA was increased in dissociated cell cultures, in ganglion explants and in vivo after axotomy. Our results suggest that CDF/LIF released from ganglionic nonneuronal cells plays an important role in regulating VIP after axotomy. 1994 John Wiley & Sons, Inc. 相似文献
65.
流行性出血热免疫球蛋白的研制 总被引:1,自引:0,他引:1
陈冬星 《微生物学免疫学进展》1994,22(3):1-6
本文首次报告采用纯化灭活流行性出血热(EHF)Ⅰ型疫苗免疫健康献血员,采集EHF抗体高滴度的血浆,用低温乙醇法及盐析法分离纯化三批EHF免疫球蛋白。结果表明:(1)采用0,1,3,4(月)或0,1,4,5(月)免疫程序,疫苗剂量1~2ml(0.15~0.30mg蛋白),受免献血员血清平均抗体滴度可达1∶406(ELISA)或1∶112(RPHI)。(2)通过生化检定,三批制品的电泳纯度为97.05%,96.84%,99.26%;IgG单体和二聚体含量为89.55%,91.30%和98.21%。(3)用空斑抑制中和试验及免疫印染试验证明所纯化的免疫球蛋白具有抗EHF病毒特异性。(4)三批EHF免疫球蛋白的效价测定,结果为ELISA滴度≥1∶512,RPHI滴度≥1∶1024,PRNT(中和抗体)滴度1∶40。按16%蛋白计,EHF免疫球蛋白的效价可达原料血浆的10倍以上。(5)无菌、安全、毒性及热原质试验检定结果,全部通过《中国生物制品规程》要求。 相似文献
66.
用共振拉曼和紫外-可见吸收光谱研究了水溶金属卟啉4-N-乙酸乙酯基-吡啶基铜卟啉和镍卟啉[简称Cu(NEAE)和Ni(NEAE)]及4-N-乙腈基-吡啶基铜卟啉[简称Cu(NACN)]与小牛胸腺DNA的相互作用。分析表明Cu(NEAE)、Ni(NEAE)和Cu(NACN)分别以外部键联、部分插入和沟槽方式与DNA作用;卟啉插入DNA时吡啶基团向卟啉环平面转动但不可能转成与之共面;而以非插入方式作用时吡啶基团会向垂直于或者平行于卟啉环平面转动。吡啶取代基的大小和空间位阻是影响相互作用方式的关键因素之一。 相似文献
67.
Depression of Neuronal Protein Synthesis Initiation by Protein Tyrosine Kinase Inhibitors 总被引:2,自引:1,他引:1
Abstract— Growth factors stimulate cellular protein synthesis, but the intracellular signaling mechanisms that regulate initiation of mRNA translation in neurons have not been clarified. A rate-limiting step in the initiation of protein synthesis is the formation of the ternary complex among GTP, eukaryotic initiation factor 2 (elF-2), and the initiator tRNA. Here we report that genistein, a specific tyrosine kinase inhibitor, decreases tyrosine kinase activity and the content of phosphotyrosine proteins in cultured primary cortical neurons. Genistein inhibits protein synthesis by >80% in a dose-dependent manner (10–80 μg/ml) and concurrently decreases ternary complex formation by 60%. At the doses investigated, genistein depresses tyrosine kinase activity and concomitantly stimulates PKC activity. We propose that a protein tyrosine kinase participates in the initiation of protein synthesis in neurons, by affecting the activity of elF-2 directly or through a protein kinase cascade. 相似文献
68.
Eviostachya hoegii Stockmans在中国五通组的首次发现 总被引:2,自引:0,他引:2
首次描述了Eviostachya hoegii Stockmans的营养部分,通过大量标本的观察,修订了前人有关其生殖部分和解剖部分的描述,并对其生殖部分进行了复原.同意Emberger(1968)的观点,将其归于Eviostachyrales中. 相似文献
69.
Guang Yi Zhang Jerry H. Wang Rajendra K. Sharma 《Molecular and cellular biochemistry》1993,122(2):159-169
Bovine brain contains two calmodulin-dependent phosphodiesterase kinases which are separated on Sephacryl S-300 column. One of these kinases has been purified to homogeneity and shown to belong to the calmodulin-dependent protein kinase II family. Phosphorylation of the 63 kDa phosphodiesterase by this purified protein kinase results in the incorporation of 1.0 mol phosphate per mol subunit and an accompanying increase in Ca2+ concentrations required for the phosphodiesterase activation by calmodulin. The protein kinase undergoes autophosphorylation to incorporate 1.0 mol phosphate per mol of subunit of the enzyme and the autophosphorylated enzyme is active, independent of the presence of Ca2+. The autophosphorylation reaction as well as the protein kinase reaction are rendered Ca2+ independent in less than 15 seconds when approximately one mol phosphate per mol protein kinase is incorporated. The result suggests that activation of phosphodiesterase phosphorylation reaction may occur prior to the activation of phosphodiesterase and phosphatase during a cell Ca2+ flux via the protein kinase autophosphorylation mechanism.Abbreviations SDS
sodium dodecyl sulfate
- EGTA
ethylene glycol bis (-aminoethyl ether)
- N,N,N,N
tetra acetic acid
- EDTA
ethylenediamine-tetraacetic acid
- cAMP
cyclic adenosine 35 monophosphate
This work is supported by grants from the Medical Research Council of Canada (JHW), the Heart and Stroke Foundation of Alberta (JHW and RKS) and the Heart and Stroke Foundation of Saskatchewan (RKS) 相似文献
70.
本文报导了薄层层析法及高压液相色谱法定性定量地测定植物油甘油酯的组成,对不同来源的脂肪酶对植物油的水解进行了研究。实验结果表明,不同来源的脂肪酶对植物油(橄榄油)的水解性不同,同一脂肪酶水解不同种类的植物油,脂肪酶的水解率也不同,脂肪酶水解植物油有最适pH和最适温度。 相似文献