共查询到18条相似文献,搜索用时 78 毫秒
1.
根据美洲商陆种子中一种抗真菌蛋白PAFP的N端部分氨基酸顺序,设计,合成一条5‘端寡核苷酸引物,通过3’-RACE技术从种子总RNA扩增出一约350bp的cDNA片段,成功地克隆到pGEM-T载体系统中。序列分析该,cDNA含有114bp的编码区,由此推导的38个氨基酸序列有36个与PAFP的序列相同,仅在第35,36位氨基酸分别由Cys,Lys代了后者的Gln和Ile。 相似文献
2.
杜仲皮中抗真菌蛋白的分离和特性研究 总被引:20,自引:0,他引:20
从杜仲(EucommiaulmoidesOliv)皮中分离纯化到一种新的抗真菌蛋白(EucommiaAntifungalProtein),简称EAFP。将切碎的杜仲树皮用NaCl溶液在组织捣碎器中高速匀浆提取,经硫酸铁沉淀、CM一cellulose离子交换层析和Bio一gel一p一10凝胶层析纯化。如此纯化而得的EAFP在SDS-PAGE胶片上显示一条分子量为5.0kD的单一带。反相HPLC显示4样品是纯的。等电聚焦电泳测得其pI值大于11.0。经还原和氨酰羧甲基化处理,用HPLC层析证明EAFP为单链。用酚一硫酸法未测出其含糖。EAFP是简单单链蛋白。氨基酸组成分析结果表明它宫含Arg、Cys和Gly,不含Met,Lys,Trp,His和Phe。未经热变性或SDS处理的EAFP与考马氏亮蓝试剂不发生显色反应。手工Edman降解法测得N一端3个氨基酸残基的顺序为Gly一Asp一Ala。用羧肽酶A和B酶解EAFP测得其C一末端为Val.0.3mg/mL蛋白明显抑制木霉、小麦赤霉菌、烟草黑茎病菌和棉花枯萎病菌菌丝的生长。蛋白在沸水中保温30min活性不变。 相似文献
3.
天麻球茎中一种抗真菌蛋白的分离和部分特性 总被引:17,自引:1,他引:17
白天麻(Gastrodia elata)顶生球茎中分离并纯化了一种抗真菌蛋白(Gastrodia Antifungal Protein),简称GAFP。在马铃薯葡糖琼脂培养基上,4μg GAFP滴加在直径0.6 cm圆纸片上可明显抑制木霉(Trichoderma reesei)菌丝生长。从每千克鲜球茎中可分离得GAFP约20 mg。用SDS-PAGE和凝胶过滤层析测得该蛋白为单多肽链,分子量14.0kD;用离子交换结合法测得电点为8.1;富含Asn,Ala,Gly和Leu,但无Met,Cys和Pro。未经热变性或SDS处理的GAFP与考马氏亮蓝试剂不发生显色反应。GAFP不具有几丁质酶和β-1,3-葡聚糖酶活性。球茎外层组织富含这种蛋白,内层薄壁组织无此蛋白。认为GAFP在阻止真菌侵染当年生顶生和侧生球茎的防卫机制中起重要作用。 相似文献
4.
美洲商陆中新发现的一种抗菌蛋白基因的克隆和表达 总被引:5,自引:0,他引:5
自美洲商陆(PhytolaccaamericanaL.)的种子中发现了一种具有38个氨基酸残基的蛋白(PaAFP),它有明显抑制立枯丝核菌(RhizoctoniasolaniKiihn)作用。依此蛋白的氨基酸序列合成引物,从该种子的mRNA中,通过逆转录PCR,获得了这一基因的具有65个氨基酸的前体蛋白cDNA序列(已在GenBank注册),将这一成熟蛋白的cDNA克隆在pGEX4T1上,并转化到大肠杆菌(E.coli),融合蛋白被大量表达。表达产物经谷胱甘肽Sepharose4B亲合层析,融合蛋白被纯化。纯化后的融合蛋白经凝血酶(thrombin)作用,将谷胱甘肽S转移酶降解,从而获得这一抗真菌蛋白 相似文献
5.
小黑麦抗真菌蛋白组分的分离纯化和性质研究 总被引:3,自引:0,他引:3
以木霉为指示菌,小黑麦中饲237种子中的蛋白提取物经过分离纯化后,得到了3种主要的抗真菌蛋白组分,经酶活检测鉴定,分别是分子量为30.5 kD的ClassⅡ型几丁质酶,两种分子量为51kD和23 kD的β-1,3-葡聚糖酶。其中几丁质酶的最适反应pH为6.0,最适反应温度为37℃,测定的N末端氨基酸序列与大麦几丁质酶的有很高的同源性。在一定条件下,这3种蛋白组分都有较强的抗木霉活性,并且有明显的协同作用,同时它们对离体易感小麦叶片上白粉菌有很好的生长抑制作用。 相似文献
6.
7.
8.
利用发根农杆菌菌株Ar1334与美洲商陆(Phytolacca americana)叶片外植体共培养转化体系,共获得58个发状根无性系(SL-1~58).以发根农杆菌Ri质粒TL-DNA上的rol C基因设计特异引物,对发状根进行PCR检测,得到了预期的560 bp目的片段,表明Ri质粒T-DNA整合到发状根基因组中.将筛选出的株系SL-7接种在MS培养基上分别置于光、暗条件下进行培养.结果发现:SL-7在暗培养条件下呈乳白色,具有多分枝、多根毛、无向地性等典型的发状根特性;在光培养条件下,发根呈粉红色,少分支且生长缓慢;以商陆抗病毒蛋白(pokeweed antiviral protein,PAP)cDNA片段为探针,分别对光、暗条件下的发状根进行Northern blot检测,发现光对PAP基因的转录具有一定抑制作用;将发状根粗蛋白提取液与TMV病毒液混合后,摩擦接种于心叶烟(Nicotiana glutinosa)离体叶片,发现暗培养的发状根粗蛋白提取液对TMV抗性明显提高.表明商陆发状根的生长及PAP基因的表达都受到光的负向调控.该结果为商陆发状根的规模化培养和PAP蛋白的离体合成优化体系的建立奠定了基础. 相似文献
9.
棕榈花序用盐溶液提取,硫酸铵沉淀,透析,DEAE—Cellulose DE—52离子交换,超滤,再经 Ultrostl AcA—44柱过滤,上 Bio—Gel P—6柱后,分离到一个相对分子量为25.6 kD的蛋白质(TP—1)。TP—1在浓度低于1.0mg/ml即可明显抑制绿色木霉菌丝的生长,对赤霉菌、棉花枯萎菌及稻瘟菌都有抑制作用。TP—1无几丁酶和β—1,3—葡聚糖酶活性.也无血球凝集活性。但经蒽酮法及PAS反应证明是一个糖蛋白。氨基酸成分分析表明,TP—1富含A_(sD)、Gly、Ala和Leu。 相似文献
10.
11.
Lu Gao Kejian Peng Yan Xia Guiping Wang Liyuan Niu Chunlan Lian 《International journal of phytoremediation》2013,15(4):307-319
Phytolacca americana L. can accumulate large amounts of heavy metals in its aerial tissues, especially cadmium (Cd) and manganese (Mn). It has great potential for use in phytoextraction of metals from multi-metal-contaminated soils. This study was conducted to further investigate the Cd- and Mn-tolerance strategies of this plant. Concentrations of non-protein thiols (NPTs) and phytochelatins (PCs) in leaves and roots increased significantly as the concentration of Cd in solution increased. The molar ratios of PCs:soluble Cd ranged from 1.8 to 3.6 in roots and 8.1 to 31.6 in leaves, suggesting that the cellular response involving PC synthesis was sufficient to complex Cd ions in the cytosol, especially that of leaves. In contrast, excess Mn treatments did not result in a significant increase in NPT or PC concentrations in leaves or roots. Oxalic acid concentrations in leaves of plants exposed to 2 or 20 mM Mn reached 69.4 to 89.3 mg (0.771 to 0.992 mmol) g–1 dry weight, respectively, which was approximately 3.7- to 8.6-fold higher than the Mn level in the 0.6 M HCl extract. Thus, oxalic acid may play an important role in the detoxification of Mn. 相似文献
12.
商陆种子抗病毒蛋白的制备及毒性分析 总被引:1,自引:0,他引:1
采用一种改进的方法,从商陆属两种植物——商陆(Phytolacca acinosa)和美州商陆(P. americana)种子中分别制备抗病毒蛋白(Pokeweed antiviral protein from seeds,简称PAP-s)。研究证明,两种植物中的PAP-s性质完全相同。经SDS-PAGE和凝胶过滤层析测得该蛋白质为单一肽链,分子量约30kD;聚丙烯酰胺等电聚焦电泳测得等电点为8.4;经Edman降解,测得N-末端氨基酸为Ile.PAP-s在兔网织红细胞裂解液中抑制蛋白质合成ID_(50)为2.6ng/ml(8.6×10~(-11)mol/1);将该蛋白质与脊髓灰质炎(?)型病毒混合接种于猴肾细胞培养液,显示出较强的抗病毒活性,浓度为4.17×10~(-8)mol/1时能抑制98%的病毒增殖。将PAP-s通过异型双功能连接剂SPDP与抗人T细胞单克隆抗体Wu71偶联制备免疫毒素,对T淋巴细胞白血病CEM细胞显示特异杀伤作用,在10~(-9)mol/1时可杀灭76.4%的靶细胞,~(14)C-亮氨酸掺入蛋白质合成抑制试验显示ID_(50)为10~(-10)—10~(-9)mol/l。 相似文献
13.
Maurice E. Edwards Elizabeth M. Harris Frederic H. Wagner M. Catherine Cross Gregory S. Miller 《American journal of botany》1988,75(12):1794-1802
Seeds of pokeweed (Phytolacca americana L.), an underinvestigated species, were studied to determine optimal conditions for laboratory germination. Soaking seeds in acid solutions prior to sowing increased rates of germination. Concentrated (conc) H2SO4 was more effective than conc NO3 or conc HCl acids. Physiological evidence from seed germination studies suggests that autotoxicity, or intraspecific inhibition, exists in pokeweed, a species known to possess several biologically active compounds. Seed germination was investigated in the laboratory with aqueous extracts of vegetative and reproductive structures of the plant. The presence of extracts from most plant parts correlated with reduced or no germination by seeds of its own species, whereas the presence of distilled water correlated with high percentages of germination by control seeds. Whether diluted with water by 5-fold (20% v/v) or undiluted, juice of pokeweed fruits completely inhibited the laboratory germination of pokeweed seeds. Also, extracts of freshly harvested mature leaves, stems, and immature fruits inhibited seed germination. However, results with root extracts, obtained from a single individual, depended more on concentration, since the highest concentration (50% v/v) inhibited germination, and lower concentrations (10 and 20% v/v) increased germination percentages over control samples. Results with extracts of juvenile leaves correlated with neither inhibition nor promotion of germination. Thus, except for juvenile leaves and the root, most extracts of the pokeweed plant inhibited seed germination with more mature structures exerting more inhibition and less mature structures exerting less or no inhibition. The ecological implication of autotoxicity is that seeds are more dispersed through time and space. In regard to seed germination by other species, especially those taxa known to possess biologically active compounds, these and other results suggest that the phenomenon of autotoxicity might be more widespread than previously suspected. 相似文献
14.
中国商陆属植物的校订 总被引:3,自引:0,他引:3
校订了我国商陆属植物,计有4种,Phytolacca clavigera W.W.Smith是多雄蕊商陆的异名,Ph.hunanensis Hand.-Mazz和Ph.zhejiangensis W.T.Fan是日本商陆的异名。Ph.esquirolii Levl.不是本属植物,而是崖豆藤野桐(大戟科)。 相似文献
15.
16.
17.
以玉米(Zea mays L.)根为材料,采用分级离心和两相分配法制备高纯度的质膜。实验比较了Tri-ton X-100和丙酮两种增溶剂对膜上ABA结合蛋白(ABA-BPm)的增溶效果。结果表明0.2%(W/V) TritonX-100的增溶效率超过85%,而丙酮法增溶效率仅达65%。放射配基(~3H-ABA)结合分析表明,增溶的膜蛋白与ABA的结合反应具有竞争性、饱和性、专一性和高亲和性等特点,而相同反应条件下的BSA则没有这些特征,证明了质膜上存在着ABA结合蛋白。实验还比较了ABA-BPm与增溶的ABA结合蛋白(ABA-BPs)与ABA的特异结合活性,结果显示ABA-BPs对反应温度和介质pH更为敏感,保持最大结合活性的时间也较短(<30min),暗示着增溶膜蛋白离开膜脂环境后更不稳定、易失活。 相似文献