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991.
天花粉蛋白Y14F/R22L定点突变及其活性研究 总被引:1,自引:0,他引:1
利用多聚酶链式反应(PCR)技术,对天然天花粉蛋白(nTCS)基因在Tyr14和Arg22两个保守残基处同时进行定点突变,即Tyr14变成Phe,Arg22变成Leu,然后克隆到pET-8c高效表达载体上,构建成重组质粒pETY14F/R22L.经序列分析,定点突变的结果与预先设计的完全一致,突变后的天花粉蛋白命名为Y14F/R22LTCS.将pETY14F/R22L转化到E.coliBL21(DE3,pLysS)中,进行表达.经CM-SepharoseCL-6B柱纯化,SDS-PAGE鉴定,纯度可达90%.RIP活性测定显示,Y14F/R22LTCS的活性比nTCS降低了7.5倍,活性变化不显著,因此,TCS的Try14和Arg22对维持其活性部位构象并不是必需的.但由于Y14F/R22LTCS在E.coli中的表达量与nTCS相比明显下降,因此,Tyr14和Arg22可能与TCS翻译后的折叠有关. 相似文献
992.
维甲酸对鼻咽癌细胞生长、表型和瘤基因表达的作用 总被引:1,自引:0,他引:1
研究了维甲酸(RA)对鼻咽癌细胞生长、表型和癌基因表达的作用.用RA诱导鼻咽癌细胞,绘制诱导前后的细胞曲线,观察细胞形态,并用Northern杂交和DNaseⅠ超敏感区分析法检测基因表达和调控.结果表明,RA能显著抑制鼻咽癌细胞的生长,前5d下降约50%.RA处理后的细胞从典型的多边形形态变成扁平、细长,类似纤维细胞状的形态.RA诱导前c-myc基因和c-Ha-ras基因HNE2细胞中高表达,而诱导后c-myc基因表达水平急剧下降,c-Ha-ras基因无明显改变.在实验中还发现RA诱导前后的c-myc基因和c-Ha-ras基因中一些重要的超敏感位点和它们的功能.由实验结果可得到如下结论:RA能促进鼻咽癌细胞分化,通过对染色体上调控位点的作用来抑制c-myc基因的表达,DNaseⅠ超敏感位点与细胞的分化程度、细胞的组织特异性和基因表达状态有关,c-myc基因可通过不同的调控方式而失活. 相似文献
993.
真核细胞基因组DNA的甲基化主要发生在CpG二核苷酸对的胞嘧啶环上(m5C)[1,2].在真核细胞基因组DNA内,约70%的CpG位点发生了甲基化.CpG位点在基因组DNA内并不是均匀分布的,多数聚集在一些基因的5’端.大量的实验证据表明,真核基因5... 相似文献
994.
本研究以兔为实验材料,对细胞核移植过程中显微操作、电融合、电活化以及移核胚的培养等基本问题进行了研究。对兔进行PMSGhCG超数排卵,收集成熟卵母细胞和16细胞胚;后者经胰蛋白酶消化,去除胶膜和透明带,在不含Ca2+、Mg2+的分离液中分成单个卵裂球;然后,分别对两者做CB预处理;首次尝试采用Wiladsen法,去除卵母细胞核、并将单个卵裂球注入透明带,同时、与McGrathSolter法进行比较;通过电融合使供体核进入去核的卵母细胞内;将所得移核胚在体外或在中间受体内培养并观察。结果表明:一、Wiladsen法与McGrathSolter法比较,核移植操作的成功率及以后的电融合率均无明显差异(Tab.1)。相对于后者,Wiladsen法更简便、易于掌握并提高去核率。二、hCG超排注射后13~15h,观察卵母细胞发现:其中,678%保留有第一极体。此时的卵子若去除1/3胞质量,去核率可以达到583%。若推迟去核时间,笫一极体退化,失去去核标志。三、比较不同电脉冲条件,发现强度为063kv/cm,持续160μs的一次电脉冲可获较高移核胚的融合率(70.8%)(Tab.2);并可使611%的成熟 相似文献
995.
Abstract Based on field population sampling of Aphis gossypii on cotton seedlings in Quzhou Experiment Station of China Agricultural University in Hebei Province in 1991, we obtained a data set consisting of 24 estimates of mean aphid density ( m , number of aphids per plant), variance (s2 ) and the proportion of plants (PT ) with no more than T aphids (T=0, 1, 2,…, 8, 10, 15, 20, respectively and defined as tally threshold). Taylor's power law fitted the data well (r2 = 0. 958). The resulting slope (1. 515) was significantly greater than 1, indicating that the spatial distribution of this aphid was in aggregated pattern. An empirical relationship between m and Pr was developed for each T value using the parameters from the linear regression In( m )= a +bln[- ln( PT )}]. The importance of the T values in reduction of sampling errors and their application to binomial sampling plans are discussed. Small T values, particularly aphid-free plant (T = 0, conventional binomial sample), could lead to spurious estimates of m from PT . A value of T from 10 to 15 was recommended to develop binomial sampling plans for the aphids on cotton seedlings because of the relatively small sampling errors. 相似文献
996.
短柄五加大,小孢子发生和雌,雄配子体发育的研究 总被引:6,自引:2,他引:4
短柄五加花药5枚,每个花药四个花粉囊。小孢子母细胞减数分裂时,胞质分裂为同时型,产生正四面体形的四分体。花药壁由表皮、药室内壁、中层和绒毡层四层细胞组成,其发育类型为双子叶型。腺质绒毡层,其细胞为二核。三细胞型花粉。子房5室,每室两个胚珠,上胚珠败育,下胚珠可育。下胚珠倒生,具单珠被,厚珠心。大孢子母细胞减数分裂形成线性排列的四个大孢子,雌配子体发育属蓼型。开花当天,花粉散开,雌配子体尚未成熟,处 相似文献
997.
NifL, an antagonistic regulator of NifA interacting with NifA 总被引:3,自引:0,他引:3
998.
Gao Yuansheng; Tolsa Jean-Francois; Shen Hai; Raj J. Usha 《Journal of applied physiology》1998,84(1):13-18
Gao, Yuansheng, Jean-François Tolsa, Hai Shen, and J. Usha Raj. Effect of selective phosphodiesteraseinhibitors on response of ovine pulmonary arteries to prostaglandinE2. J. Appl. Physiol. 84(1): 13-18, 1998.Several adenosine3,5-cyclic monophosphate (cAMP)-hydrolyzingphosphodiesterase isozymes are present in the pulmonary vasculature.The present study was designed to determine the effect of selectiveinhibitors of phosphodiesterase subtypes on prostaglandinE2(PGE2)-induced relaxation ofisolated fourth- generation pulmonary arteries of newborn lambs.PGE2 and forskolin causedpulmonary arteries to relax and induced an increase in theintracellular cAMP content in the vessels. The relaxation and change incAMP content were augmented by milrinone and rolipram, inhibitors ofphosphodiesterase type 3 (PDE3) and type 4 (PDE4), respectively. Theaugmentation in relaxation and the increase in cAMP content caused bymilrinone plus rolipram was greater than the sum of theresponses caused by either of the inhibitors alone.8-Methoxymethyl-1-methyl-3-(2-methylpropyl)xanthine, an inhibitor of phosphodiesterase type 1, had no effect on relaxation andchange in cAMP induced by PGE2 andforskolin. Acetylcholine alone had no effect on cAMP content in thevessels but augmented the relaxation and the increase in cAMP inducedby PGE2 and forskolin in arterieswith endothelium. This effect was not observed in arteries withoutendothelium or in arteries with endothelium treated withNG-nitro-L-arginine.These results suggest that PDE3 and PDE4 are the primary enzymeshydrolyzing cAMP of pulmonary arteries of newborn lambs and that aninhibition of both PDE3 and PDE4 would result in a greater effect thanthat caused by inhibition of either one of the subtype isozymes alone.Furthermore, endothelium-derived nitric oxide may enhance cAMP-mediatedrelaxation by inhibition of PDE3. 相似文献
999.
Trichosanthin(TCS)isanimportantmemberofribosomeinactivatingproteins[1].ItpossessesNglycosidaseactivityremovingadenine(ADE)atpositionA4324of28SrRNA[2].TheactivepocketofNglycosidasehasbeenestablishedthroughthecrystalstructuresofTCS,αMMCandricinandassayofmutants… 相似文献
1000.
The relation between the fast phase of ms-DLE (delayed light emission measured with a phosphoroscope) and the proton released from water oxidation in spinach chloroplasts was studied in several aspects. When photophosphorylation was allowed to be coupled to the Hill reaction the intensity of the fast phase of ms-DLE of chloroplast was lowered more at 1 °C than at 25 °C, and the photophosphorylation rate within 40 ms of flashing light was higher at 1 °C than at 25 °C. Adding the subunit of ATP synthase to the chloroplast preparation to block the leakage of protons through ATP synthase, the intensity of the fast phase of ms-DLE was enhanced, to a larger extent at 1 °C than at 25°C. When the ms-DLE was measured under isotonic conditions, the intensity of fast phase of ms-DLE enhanced by proton released from oxidation of water was more pronounced. The above results support the suggestion that under lower temperature and isotonic conditions, the proton released from water oxidation was liable to be localized and could enhance the intensity of the fast phase of ms-DLE more effectively. 相似文献