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91.
利用RT-PCR技术分离低密度脂蛋白受体基因cDNA,将长为2605bp的cDNA插入质粒pJN6,并经全序列测定证实,该序列与已报道序列相比,存在两个变异碱基,即第754位C→T,和1654位的G→A,这两个变异碱基并不改变编码的氨基酸,利用LDL受体基因cDNA中的ClaⅠ片段作为探针,检测到LDL受体基因上外显子8的StuⅠ位点是个限制性片段长度多态性位点。所克隆的cDNA含有可译框架的全部密码,因此可作为基因表达材料。 相似文献
92.
水稻巯基蛋白酶抑制剂(CPI)经用二硫苏糖醇,对氯汞苯甲酸和碘乙酸修饰后,对木瓜蛋白酶的抑制活性并无改变;用N-乙基顺丁烯二酰亚胺与CPI反应,可以测出CPI分子内有19个巯基被修饰,被修饰后,抑制活性仍无改变,表明水稻CPI的抑制活性不需要巯基参与;应用N-溴代丁二酰亚胺与CPI反应,可测出CPI分子内有2个Trp被修饰,修饰后,抑制活性全部丧失,表明Trp是保持抑制活性所必需的基团。水稻巯基蛋白酶抑制剂和丝氨酸蛋白酶抑制剂对稻瘟病菌丝体的生长均有抑制作用,但后者的抑制作用比前者更强,若将两种抑制剂混合使用,则对稻瘟病菌丝体的抑制作用非常强烈;当抑制剂加入量达72μg时,即可产生明显的抑制作用。 相似文献
93.
Yuh-Shyong Yang A.David Marshall Peter Mcphie Wei-Xi Athena Guo Xiaofu Xie Xiang Chen William B. Jakoby 《Protein expression and purification》1996,8(4):423-429
A phenol sulfotransferase from rat liver (EC 2.8.2.9), expressed inEscherichia colifrom a single cDNA, was purified as two separable but catalytically active proteins. The proteins appeared to be identical to each other and to the natural liver sulfotransferase by comparison of their amino acid constitution, amino-terminal end group, and interaction with a polyclonal antibody raised against the liver enzyme. Each of the recombinant forms, α and β, catalyzed the sulfuryl group transfer from 4-nitrophenylsulfate to an acceptor phenol, a reaction in which 3′-phospho-adenosine 5′-phosphate (PAP) is a necessary intermediate. Only form β, however, catalyzes the physiological transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulfate (PAPS) to the free phenol. Evidence is presented that sulfotransferase α, but not β, has 1 mol of PAP tightly bound per enzyme dimer. The ability to utilize PAPS as a sulfate donor could be altered: form α could be treated and purified as form β to acquire the ability to use PAPS, whereas form β was treated by extended incubation with PAP, lost its ability to use PAPS, and was purified as form α. 相似文献
94.
Fate of the human immunodeficiency virus type 1 provirus in infected cells: a role for vpr. 总被引:11,自引:11,他引:0 下载免费PDF全文
V Planelles F Bachelerie J B Jowett A Haislip Y Xie P Banooni T Masuda I S Chen 《Journal of virology》1995,69(9):5883-5889
We investigated the fate of human immunodeficiency virus type 1 (HIV-1) viral DNA in infected peripheral blood lymphocytes and immortalized T-cell lines by using a replication-defective HIV-1. We observed that integrated HIV-1 DNA and viral gene expression decrease over time. A frameshift mutation in vpr resulted in maintenance of the HIV-1 provirus and stable persistence of viral expression. Transfection of vpr together with the neomycin resistance gene in the absence of other viral genes decreased the formation of geneticin-resistant colonies, indicating either a cytotoxic or a cytostatic effect upon cells. Therefore, maintenance of HIV-1 infection within an infected proliferating population is due to two competing processes, the rate of viral spread and the degree of cell growth inhibition and/or death induced by Vpr. 相似文献
95.
Identification and analysis of a retinoblastoma binding motif in the replication protein of a plant DNA virus: requirement for efficient viral DNA replication. 总被引:22,自引:3,他引:19 下载免费PDF全文
Geminiviruses are plant DNA viruses with small genomes whose replication, except for the viral replication protein (Rep), depends on host proteins and, in this respect, are analogous to animal DNA tumor viruses, e.g. SV40. The mechanism by which these animal viruses create a cellular environment permissive for viral DNA replication involves the binding of a virally encoded oncoprotein, through its LXCXE motif, to the retinoblastoma protein (Rb). We have identified such a LXCXE motif in the Rep protein of wheat dwarf geminivirus (WDV) and we show its functional importance during viral DNA replication. Using a yeast two-hybrid system we have demonstrated that WDV Rep forms stable complexes with p130Rbr2, a member of the Rb family of proteins, and single amino acid changes within the LXCXE motif abolish the ability of WDV Rep to bind to p130Rbr2. The LXCXE motif is conserved in other members of the same geminivirus subgroup. The presence of an intact Rb binding motif is required for efficient WDV DNA replication in cultured wheat cells, strongly suggesting that one of the functions of WDV Rep may be the linking between viral and cellular DNA replication cycles. Our results point to the existence of a Rb-like protein(s) in plant cells playing regulatory roles during the cell cycle. 相似文献
96.
A chalcone synthase-like cDNA from rice anther 总被引:3,自引:0,他引:3
97.
以细胞形态观察、细胞存活率、乳酸脱氢酶释放和前列环素测定等方面研究载脂蛋白AⅡ保护内皮细胞的作用.结果显示,载脂蛋白AⅡ可部分拮抗低密度脂蛋白对内皮细胞的损伤作用, 维持内皮细胞形态基本正常,乳酸脱氢酶释放减少,前列环素合成增加. 相似文献
98.
A Mitosis-specific Phosphorylation of the Gap Junction Protein Connexin43 in Human Vascular Cells: Biochemical Characterization and Localization 总被引:3,自引:0,他引:3 下载免费PDF全文
Han-qing Xie Dale W. Laird Tsg-Hui Chang Valerie W. Hu 《The Journal of cell biology》1997,137(1):203-210
Western blotting studies revealed that connexin43 (Cx43), one of the major gap junction proteins in human vascular endothelial cells, is posttranslationally modified during mitosis. This mitosis-specific modification results in a Cx43 species that migrates as a single protein band and was designated Cx43m. Cx43m was shown to be the result of additional Ser/Thr phosphorylation as indicated by: (a) the increased gel mobility induced by both alkaline phosphatase and the Ser/ Thr-specific protein phosphatase-2A (PP2A) and (b) the removal of virtually all 32Pi from Cx43m by PP2A. Immunofluorescent confocal microscopy of mitotic cells revealed that Cx43 is intracellularly located, while in nonmitotic cells Cx43 is located at regions of cell–cell contact. Dye coupling studies revealed that mitotic endothelial cells were uncoupled from each other and from nonmitotic cells. After cytokinesis, sister cells resumed cell coupling independent of de novo protein synthesis. The mitosis-specific phosphorylation of Cx43 correlates with the transient loss of gap junction intercellular communication and redistribution of Cx43, suggesting that a protein kinase that regulates gap junctions is active in M-phase. 相似文献
99.
Sato Yoji; Weil Max Harry; Tang Wanchun; Sun Shijie; Xie Jianlin; Bisera Joe; Hosaka Hidehiro 《Journal of applied physiology》1997,82(2):558-562
Sato, Yoji, Max Harry Weil, Wanchun Tang, Shijie Sun,Jianlin Xie, Joe Bisera, and Hidehiro Hosaka. EsophagealPCO2 as a monitor of perfusionfailure during hemorrhagic shock. J. Appl.Physiol. 82(2): 558-562, 1997.Measurement ofgastric wall PCO2(PgCO2) bytonometric method has emerged as an attractive option for estimatingvisceral perfusion during circulatory shock. However, gastric acidsecretion obfuscates the tonometric measurement. We, therefore,investigated the option of measuringPCO2 in the esophagus to minimizethese restraints. Hemorrhagic shock was induced in five Sprague-Dawleyrats, and five rats served as sham controls.PgCO2 wasmeasured with an ion-sensitive field effect transistor that wassurgically implanted into the gastric wall. Esophageal luminalPCO2(PeCO2) wasmeasured by a second ion-sensitive field effect transistor sensor.During hemorrhagic shock, mean aortic pressure declined from 150 to 50 mmHg. Gastric blood flow decreased from 58 to 12 ml · min1 · 100 g1 (21% of preshock) andesophageal blood flow from 44 to 7 ml · min1 · 100 g1 (16% of preshock).PgCO2simultaneously increased from 47 to 116 Torr andPeCO2 from 47 to 127 Torr. The increases inPgCO2 werehighly correlated with increases inPeCO2(r = 0.90). Esophageal tonometry may,therefore, serve as a practical alternative to gastric tonometry. 相似文献
100.
Xue Bin Zheng Misao Tsubokura Yongkun Wang Chun Xie Tetsuji Nagano Kazuhiko Someya Tomoko Kiyohara Kiyomi Suzuki Takeshi Sanekata 《Microbiology and immunology》1995,39(10):821-824
Thirty strains of Yersinia pseudotuberculosis were isolated from rabbits (17 strains), wild rats (9 strains) and house rats (4 strains) in China between 1990 and 1993. The biochemical properties of these isolates were identical with those of Y. pseudotuberculosis and no special characteristics were found in these strains. Serologically, serogroups 4b and 5b were identical to isolates found in Japan, and a new serogroup 1c and unclassified strains have also been detected. The existence of virulence-associated properties were different among strains. The pYV plasmid was detected from 6 strains of 30 isolates. This report documents the presence of Y. pseudotuberculosis in China, providing important epidemiological information. 相似文献