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281.
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
282.
用化学修饰、内源荧光和荧光淬灭等方法研究了油麻藤凝集素(MSL)的溶液构象变化和微环境的构象特征。研究发现MSL分子中总共有9个色氨酸(Trp)残基,它们的荧光能被丙烯酰胺淬灭,但不易为KI接近而淬灭,MSL经N-溴代琥珀酰亚胺(NBS)修饰后,其内源性荧光发射谱发生相应变化,结果表明MSL分子中部分Trp残基埋藏于分子内部,而位于分子表面的Trp残基可能处于分子的疏水袋中。 相似文献
283.
木瓜蛋白酶PPAⅢ自水解作用 总被引:1,自引:0,他引:1
木瓜蛋白酶PPAⅢ自水解作用王秀艳,周慧,葛玉斌,李惟(吉林大学分子生物学系,长春130023)PPAⅢ、papain、pepsine、stembromelain等都属于以Cys-SH为活性中心的酶,同种分子之间存在着一种相互切割的趋势,称为酶自水解... 相似文献
284.
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
285.
黄精凝集素Ⅱ分子稳定性与生物学活性研究鲍锦库,曾仲奎,周红(四川大学生物系,成都,610064)本文在黄精凝集素Ⅱ纯化及性质研究的基础上,应用多种变性条件,研究其分子特性,同时对分子的巯基和色氨酸进行修饰,研究该凝集素分子保持其生物学活性与这些基团的... 相似文献
286.
Hongzhang Zhou 《Insect Science》1996,3(4):329-337
Abstract The seasonality of the field population of Lagria hirta L. is remarkable: adults occur only from the end of June to the beginning of August, whereas larvae exist in the whole year except June. Based on the data of field observations and larval head-width measuring, the author demonstrated that L. hirta would reproduce in a very short interval in June to August; eggs hatch and develop (molt) into the 3rd, 4th or 5th (sometimes 6th) instar before winter; after larval overwintering, the beetles complete the larval development after total 8–10 times of molts and pupate in June. The larval development rate is decreased but not completely stopped during winter (the nonparametric median test, P<0. 01). This result changes the hitherto opinion of 4 or 5-instar larval development of L. hirta and will stimulate the studies on its life history. 相似文献
287.
Papillomavirus capsid binding and uptake by cells from different tissues and species. 总被引:22,自引:14,他引:8
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M Müller L Gissmann R J Cristiano X Y Sun I H Frazer A B Jenson A Alonso H Zentgraf J Zhou 《Journal of virology》1995,69(2):948-954
The inability of papillomaviruses (PV) to replicate in tissue culture cells has hampered the study of the PV life cycle. We investigated virus-cell interactions by the following two methods: (i) using purified bovine PV virions or human PV type 11 (HPV type 11) virus-like particles (VLP) to test the binding to eukaryotic cells and (ii) using different VLP-reporter plasmid complexes of HPV6b, HPV11 L1 or HPV11 L1/L2, and HPV16 L1 or HPV16 L1/L2 to study uptake of particles into different cell lines. Our studies showed that PV capsids bind to a broad range of cells in culture in a dose-dependent manner. Binding of PV capsids to cells can be blocked by pretreating the cells with the protease trypsin. Penetration of PV into cells was monitored by using complexes in which the purified PV capsids were physically linked to DNA containing the gene for beta-galactosidase driven by the human cytomegalovirus promoter. Expression of beta-galactosidase occurred in < 1% of the cells, and the efficiency of PV receptor-mediated gene delivery was greatly enhanced (up to 10 to 20% positive cells) by the use of a replication-defective adenovirus which promotes endosomal lysis. The data generated by this approach further confirmed the results obtained from the binding assays, showing that PV enter a wide range of cells and that these cells have all functions required for the uptake of PV. Binding and uptake of PV particles can be blocked by PV-specific antisera, and different PV particles compete for particle uptake. Our results suggest that the PV receptor is a conserved cell surface molecule(s) used by different PV and that the tropism of infection by different PV is controlled by events downstream of the initial binding and uptake. 相似文献
288.
稀有鮈鲫──一种新的鱼类毒性试验材料 总被引:7,自引:0,他引:7
本文研究了稀有鲫(Gobiocyprisrarus)作为毒性试验材料的可行性。采用换水式试验,在硬度为200mg/L(以CaCO3计)、pH7.8±0.2、温度24-25℃条件下研究了铬、铜、锌和五氯酚(PCP)对稀有鲫的急性毒性。重铬酸钾对2日龄稀有鲫的24h和96h和LC50控制范围分别263.6-334.7和1153-178.5mg/L(n=8)。铬、铜、锌和五氯酚对2日龄稀有鲫的急性毒性值(96hLC50)范围,从铜的52.2μg/L到铬的52000μg/L,毒性大小的顺序是铜>五氯酚>锌>铬。研究结果表明,稀有鲫有可能发展成为一种较为理想的毒性试验材料。 相似文献
289.
290.
Identification of a 175 kDa protein as the ligand-binding subunit of the rat liver sinusoidal endothelial cell hyaluronan receptor 总被引:2,自引:1,他引:1
The rat liver sinusoidal endothelial cell (LEC) hyaluronan (HA)receptor was previously identified using a photoaffinity HAderivative (J. BioL Chem., 267, 2045120456, 1992). Twopolypeptides with Mr = 175,000 and 166,000, were consistentlycrosslinked, suggesting that the LEC HA receptor is an oligomer.Whether one or both subunits participate in HA binding, wasnot determined. Here we investigate the HA-subunit interactionsand the potential oligomeric nature of the LEC HA receptor.When Sephacryl-400 gel filtration chromatography was used toenrich the HA receptor, the 175 kDa polypeptide was the majorband seen by SDS-PAGE analysis. Little staining was seen at166 kDa, suggesting that the 175 kDa protein could be separatedfrom the 166 kDa protein and still retain HA-binding activity.A ligand blot assay was used to determine if each individualsubunit could bind HA. LEC proteins were separated by nonreducingSDS-PAGE, and then immobilized onto nitrocellulose. 125I-HAbound to a 175 kDa polypeptide but not to the 166 kDa protein.A high molecular weight band of 相似文献