共查询到18条相似文献，搜索用时 296 毫秒
1.
通过转录组测序获得了一个广叶绣球菌(Sparassis latifolia)编码的溶血素基因(latifolysin),进行生物信息学分析,并利用实时定量PCR分析溶血素基因表达水平,采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)标记结合二维液相色谱串联质谱鉴定分析溶血素表达差异.结果表明latifolysin基因全长为399 bp,编码133个氨基酸.保守功能域搜索显示latifolysin具有与杨树菇溶血素(aegerolysins)蛋白家族相同的结构域和功能位点,两者序列相似性40％.杨树菇溶血素家族蛋白在真菌中呈不连续性分布.系统进化树结果显示绣球菌溶血素隶属于担子菌类群,与草菇(Volvariellavolvacea)遗传关系较近.实时定量PCR和蛋白定量结果表明,latifolysin和绣球菌溶血素在广叶绣球菌幼菇期表达量最高,且显著高于菌丝体中表达水平. 相似文献
2.
利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)标记结合二维液相色谱串联质谱(two-dimensional liquid chromatography tandem mass spectrometry,2D LC-MS/MS)技术,研究广叶绣球菌(Sparassis latifolia)原基期、幼菇期和成熟子实体阶段的差异表达蛋白质组.采用Q-Exactive质谱鉴定并经ProteinPilot软件搜库,对所获得的差异蛋白进行GO (gene ontology)、KEGG(kyoto encyclopedia of genes and genomes)和转录因子注释分析.结果共鉴定到可信蛋白2305个,其中2219个蛋白具有相对定量信息.与原基期相比,幼菇期显著上调蛋白104个,下调蛋白142个,子实体阶段显著上调蛋白155个,下调蛋白460个.GO分子功能提示这些差异性蛋白主要参与催化活性、蛋白结合和水解酶活性.KEGG代谢通路分析结果显示,差异蛋白主要涉及碳代谢、氨基酸合成、核糖体、糖酵解/糖衍生等代谢过程.差异蛋白中与信号传导和转录因子相关的蛋白数量分别为27个和7个. 相似文献
3.
《食用菌学报》2015,(1)
依据草菇(Volvariella volvacea)基因组和转录组数据,采用ZOOM软件将转录组Reads定位到基因组的分析方法,从草菇中鉴定了一个多糖单加氧酶基因Vv-lpmo1,对其进行生物信息学分析,并对其高效诱导物进行筛选,采用定量PCR方法研究其在降解稻草时表达量的变化规律。结果表明:草菇Vv-lpmo1基因DNA序列长1432bp,有9个内含子,预测编码305个氨基酸的蛋白质;生物信息学分析表明该蛋白质有分泌信号肽,且含有两个完整的保守结构域:N端的glycoside hydrolase family 61(GH61)结构域和C端的cellulose-binding domain(CBD)结构域。通过对葡萄糖、无碳源(饥饿诱导)、微晶纤维素、滤纸、稻草和木屑6种不同基质诱导草菇菌丝的定量PCR,结果筛选出稻草对Vv-lpmo1基因有最强的诱导作用。进一步研究稻草诱导不同时间的变化过程中,该基因的表达规律为0~4h先下调;4~12h恢复到初始水平;12h以后开始迅速上调并维持较高水平。本研究表明草菇多糖单加氧酶基因Vv lpmo1属于诱导表达型基因,可能在草菇降解稻草基质中起着重要作用。 相似文献
4.
'白核'龙眼种子败育不同时期差异蛋白分析 总被引：1，自引：0，他引：1
以'白核'龙眼种子为试验材料,比较两种蛋白质的提取方法,确定了酚抽法提取龙眼种子蛋白.应用蛋白质组学研究技术,分析了龙眼种子败育4个不同时期的蛋白质组变化,共发现21个差异蛋白,其中9个上调表达,12个下调表达.通过MALDI/TOF/TOF-MS/MS质谱分析和蛋白质数据库检索,共鉴定出12个差异蛋白,分别为胞质苹果酸脱氢酶、丙酮酸脱氢酶、磷酸甘油酸变位酶、RuBisCO大亚基结合蛋白α、β亚基、17 kD HSP、抗坏血酸过氧化物酶、胞质抗坏血酸过氧化物酶、半胱氨酸蛋白酶、Dessication-related protein以及两个未知蛋白.这些鉴定出的差异蛋白质与能量和物质代谢、分子伴侣功能、自由基清除和抗氧化作用、细胞凋亡等生理过程密切相关,可能参与了龙眼种子败育过程. 相似文献
5.
《食用菌学报》2015,(2)
以金针菇(Flammulina velutipes)子实体、初生菌丝和次生菌丝为材料,采用荧光染色法对子实体的囊状体、担子、担孢子,初生菌丝和次生菌丝、粉孢子进行镜检、观察细胞核。结果为子实体中囊状体无细胞核,只有担子和担孢子内有细胞核,细胞核分裂不同步;细胞核变化过程为:由双核经历核融合,形成单核,经过二次减数分裂和一次有丝分裂后形成8个核,以2个为一组进入担孢子,形成双核担孢子,最后留下无核的担子。初生菌丝(由担孢子萌发的菌丝)细胞核不定,为1~5个核,次生菌丝(初生菌丝细胞质融合形成的双核菌丝)细胞核为2个,初生菌丝和次生菌丝形成的粉孢子均为1~2个核,以单核为主。 相似文献
6.
桃叶片总蛋白双向电泳技术的研究 总被引：1，自引：0，他引：1
采用改进的O'Farrell双向电泳系统,研究适于桃树叶片蛋白质组分析的双向电泳方法。通过对不同蛋白样品提取方法、不同pH梯度范围、不同上样量的比较,探索出一套适合桃树叶片总蛋白分析的双向电泳方法,即以TCA/丙酮沉淀法提取叶片蛋白质样品,采用两性电解质配比pH3～10︰pH5～7为1∶4的IEF胶条,按90μg蛋白上样量,并结合适宜的双向电泳参数,可获得背景清晰、重复性较好、蛋白分辨率高的双向电泳图谱。经此方法分离的桃叶片蛋白质经串联质谱分析(MS/MS)得到了较好的鉴定。 相似文献
7.
以3年生笃斯越橘苗为材料,将苗木分别置于对照(23℃,日照长度14 h),低温短日照(4℃,日照长度10 h)和低温长日照(4℃,日照长度14 h)的人工气候室中。处理21 d后,采用同位素标记相对和绝对定量(iTRAQ)蛋白质组学技术测定枝条蛋白质表达变化情况。结果显示:通过质谱鉴定出5 972个蛋白质,600个差异表达蛋白,其中在低温短日照与对照比对组中,丰度显著上调蛋白有140个,丰度显著下调蛋白有114个;在低温长日照与对照比对组中,丰度显著上调蛋白有255个,丰度显著下调蛋白有122个;在低温短日照与低温长日照比对组中,丰度显著上调蛋白有39个,丰度显著下调蛋白有187个。这些蛋白主要参与(1)RNA代谢,(2)蛋白质翻译后修饰,(3)碳水化合物转运和代谢,(4)能量代谢和转换,(5)脂质代谢,(6)次生代谢产物合成,(7)抗氧化与胁迫防御,(8)光合作用,(9)无机离子转运和代谢。结果表明与抗冻性有关的蛋白差异表达主要受低温诱导,少数蛋白表达受低温和光周期共同影响,低温短光周期更有利于抗冻性形成。低温锻炼可显著提高RNA代谢、蛋白质翻译后修饰、抗氧化和防御反应、次生代谢产物合成以及脂质代谢过程中许多蛋白质的丰度,提示这些代谢途径和相关蛋白可能在笃斯越橘抗冻机制中起重要作用。 相似文献
8.
食用菌菌种结实性和产量性状早期鉴定的初步研究 总被引：2，自引：2，他引：0
实验以34个草菇（Volvariella volvacea）和95个香菇（Lentinula edodes）培养物~(**)为材料,进行了食用苗苗种结实性早期鉴定的研究;以16个侧耳（Pleurotus）~***和14个金针菇（Fla-mmulina velutines）菌株为材料,进行了食用菌菌种产量性状早期鉴定的研究.研究表明,草菇和香菇菌种培养物在平板上的菌落形态特征与结实性有密切关系.草菇培养物的苗落形态还与菌丝形态、长速、酯酶同工酸酶谱、可溶性蛋白带、发菌和出菇等密切相关.侧耳和金计菇菌丝的胸外羧甲基纤维素酶（CMC酶）活力与子实体产量成正相关,可以作为其子实体产量性状早期鉴定的指标之一. 相似文献
9.
《中国果树》2017,(2)
蛋白质样品制备方法是蛋白质组学分析的关键。为建立适于苹果花器官总蛋白提取及分析方法,以10年生‘华月’苹果花序为试材,通过优化提取条件,确立了适于苹果花器官总蛋白提取的技术方法。基于此,开展了苹果花器官蛋白双向电泳分析,并对随机筛选的32个蛋白点进行质谱(MALDI-TOF-TOF/MS)鉴定,经数据库检索,32个蛋白点均得到成功鉴定,按照功能划分为代谢及能量产生相关、抗性相关、蛋白质合成相关、细胞结构相关、调节相关及未知功能蛋白等6类。该研究通过优化蛋白质样品制备方法,进而完成双向电泳分析及质谱鉴定,为进一步利用苹果花器官开展苹果抗病蛋白质组学研究奠定了基础。 相似文献
10.
《食用菌学报》2020,(2)
以黄色金针菇(Flammulina filiformis)单核菌株Y33和白色金针菇单核菌株W8为供试菌株,采用双向核迁移技术,获得一对具有相同细胞核、不同细胞质的双核菌株Y8-33和W8-33(简称异质双核菌株)。采用菌落形态观察、显微镜检查与分子标记对这两个双核菌株进行验证分析,并以菌株W8-33菌丝样品为对照,采用双向凝胶电泳技术对金针菇异质双核菌株菌丝的蛋白质组进行分析。结果表明:异质双核菌株Y8-33和W8-33菌丝均具有锁状联合结构,在菌落形态上呈现明显差异,但ISSR分子标记的PCR扩增产物具有很高的相似性。在Y8-33菌丝样品中共鉴定出17个差异蛋白,包括8个上调蛋白、9个下调蛋白;其中,14-3-3蛋白、微管蛋白、谷胱甘肽-S-转移酶、果糖二磷酸醛缩酶、ARF/SAR蛋白家族以及热激蛋白HSP90、HSP70和HSS1差异蛋白可能在由于受体细胞不同所导致的不同细胞质环境下,对金针菇的生长生理起到重要的调控作用。 相似文献
11.
双孢蘑菇同核原生质体育种技术研究：Ⅱ.同核原生质体的杂交 总被引：3，自引：1，他引：2
为建立双孢蘑菇同核原生质体杂交育种体系,本文对同核原生质体亲合性反应及杂交异核体F_1代的农艺性状和F_2代的分离与变异规律进行了研究。结果表明:双孢蘑菇同核原生质体的杂交是一个较复杂的性亲合与核迁移过程,其杂交率较低。在按气生型×气生型、气生型×匍匐型、匍匐型×匍匐型组合的520个杂交配对中,共获得113个杂交异核体,平均杂交率为22%。本研究同时建立了以菌落形态、菌丝生长速度、羧甲基纤维素酶相对活性以及酯酶同功酶电泳为手段的杂交异核体鉴定体系。 相似文献
12.
13.
AIM：To screen the possible serum biomarkers of Parkinson’s disease. METHODS：The surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was used to screen the serum samples from 44 cases of Parkinson’s disease and 60 control subjects. The differentially expressed protein peaks were selected and isolated with high-performance liquid chromatography (HPLC), and processed with enzyme before analysis by liquid chromatography-mass spectrometry tandem mass spectrometry (LC-MS/MS). The data mining was performed by Xcalibur program component BioWorks 3.2. RESULTS：Three differentially expressed protein peaks were selected as potential serum biomarkers from the patients of Parkinson’s disease: the protein at 8 937 m/z peak showed significant increase (27.47±16.58 in Parkinson’s disease group, and only 5.01±3.47 in control group), and the proteins at 6 636 and 8 697 m/z peaks showed significant decreases (5.43±2.66 and 20.22±9.57, respectively, in Parkinson’s disease group, and 18.85±7.56 and 51.13±26.22, respectively, in control group). The proteins at 6 636, 8 697 and 8 937 m/z peaks were identified as apolipoprotein C-I, apolipoprotein C-III and complement 3a,respectively. Combined use of these 3 biomarkers effectively distinguished the subjects between Parkinsons disease group and control group. The detection rate of the patients with Parkinsons disease was 90.0% (27/30), and the detection rate of the healthy sibkects was 92.5% (37/40). CONCLUSION：The apolipoprotein C-I, apolipoprotein C-III and complement component 3a identified as potential markers of Parkinson’s disease have diagnostic value in clinical application. 相似文献
14.
ZHANG Jin DING Mei-ping LIU Zhao ZHANG Bao-rong Li Guo-liang ZHOU Fu-you GU Miao 《园艺学报》2006,22(9):1779-1783
AIM:To examine the expression profiles of both genes and proteins in hippocampus of rats with temporal lobe epilepsy (TLE) for revealing the molecular mechanisms of TLE and looking for the candidate targets and new therapeutic approaches in clinical practice.METHODS:Rat temporal lobe epilepsy was induced by administration of lithium chloride and pilocarpine (LiCl-PILO).The expression spectra of genes and proteins were constructed through the techniques of cDNA microarray,two-dimensional (2D) electrophoresis and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).Subsequently,the differentially expressed genes and proteins were identified and analyzed.RESULTS:There were 192 genes of differential expression observed in hippocampal tissues of LiCl-PILO-induced temporal lobe epilepsy,and 159 genes have been registered in Genbank database,in which 84 genes were up-regulated while 75 genes were down-regulated.78 protein spots of differential display were screened out,in which 31 proteins were detected to be down-regulated and 47 were up-regulated.Finally,5 proteins were identified.CONCLUSION:These genes and proteins found in our study may play pivotal roles in the pathogenic mechanisms of epilepsy and may promise new therapeutic targets for refractory epilepsy in the future. 相似文献
15.
JI Ying-hong LU Yi LI Na JIN Hong YANG Peng-yuan KONG Xiang-yin YAN Shun-sheng 《园艺学报》2009,25(7):1415-1419
AIM: To separate total lens proteins of congenital inherited cataract in mice and to observe the alteration of proteins after gene mutation.METHODS: We studied the mice with a spontaneous mutation of crystallin gamma S (Crygs) transmitted as a recessive trait. Total proteins were extracted and separated using immobilized pH gradient (IPG), two-dimensional electrophoresis (2-DE) and colloidal Coomassie brilliant blue (CBB) staining. The image analysis was carried out using PDQuest 7.30 software package. Several significantly differential proteins in gel were identified by matrix assisted laser adsorption/ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS). RESULTS: As the 882 μg sample was added, we detected (417±53) spots and (370±41)spots in cataract and normal lens, respectively. As the 190 μg sample was loaded, we detected (60±7) spots and (57±5) spots in cataract and normal lens, respectively. Seven kinds of differential proteins were identified, including BFSP/filensin, γS, γF, βA1, βB1, βB2 and αB. In crystalline lens of mutant mice, γS and beaded-filament structure protein (BFSP/filensin) were not detected. γF was down-expressed (＜4 fold) while βA1, βB1, βB2 and αB were over-expressed (＞4 fold) in mutant cataract. The latter proteins were less MW than normal, suggesting that they were possibly truncated.CONCLUSION: 2-DE and mass spectrometry can help to assess and analyze the function of proteins as a novel approach. The mutant Crygs gene can lead to the abnormity of γS crystallin, which can induce the changes of skeletonal protein (BFSP/filensin) and other crystallins (γF, βA1, βB1, βB2 and αB), and then evoke cataract secondarily. 相似文献
16.
Tuberization is a complex and multilevel developmental process. Many important metabolic changes in the early stage of tuberization are crucial to the tuber differentiation and development. In this study, we attempted to identify proteins differentially expressed in the early stage of in vitro tuberization in taro (Colocasia esculenta var. antiquorum) by two-dimensional polyacrylamide gel electrophoresis (2-DE) and mass spectrometry (MS). Protein samples from shoot tips cultured in 8% sucrose media at 0, 2, 4, 6 and 8 d were separated with 2-DE. A total of 13 differentially expressed proteins were analyzed with MS. Four proteins via, glyceraldehyde-3-phosphate dehydrogenase, alcohol dehydrogenase, chloroplast protein synthesis elongation factor (EF-Tu), and ankyrin repeat protein HBP1 were successfully identified during in vitro tuberization of taro. This implies that important metabolic changes, including sucrose metabolism, signal transduction and cell defense, occurred in the early stage of in vitro tuberization in taro. 相似文献
17.
建立了液相色谱-串联质谱仪(Liquid Chromatography-Tandem Mass Spectrometry,LC-MS/MS)同时测定糙皮侧耳(Pleurotus ostreatus)中吲哚乙酸、吲哚丙酸、吲哚丁酸、赤霉素、2,4-二氯苯氧乙酸、6-苄基腺嘌呤、4-氯苯氧乙酸、6-糠氨基嘌呤和芸苔素内酯9种植物生长调节剂残留的定量分析方法,样品经含1%(v/v)乙酸的乙腈溶液提取,于分散固相萃取管(含混匀的150mg MgSO_4和50mg C_(18))净化后用LC-MS/MS检测。结果表明:9种植物生长调节剂的峰面积与其浓度呈良好线性关系,方法最低检出限为0.5~1.0μg/kg;回收率为80.3%~96.5%,相对标准偏差为0.88%~7.31%,该方法准确、可靠,可以同时满足糙皮侧耳中9种植物生长调节剂残留检测的需要。对市场上50份糙皮侧耳的测定结果为:吲哚乙酸检出6次(2.3~96.7μg/kg)、吲哚丙酸2次(21.3、87.8μg/kg)、吲哚丁酸5次(3.4~91.2μg/kg)。 相似文献
18.
侧耳与香菇属间原生质体融合育种研究 总被引：5，自引：1，他引：4
侧耳、香菇皆属四极性异宗结合食用菌,其异核双核菌丝体均具“锁状联合”形态标记。本研究以PEG-Ca~(++)为诱导剂,以紫孢侧耳和香菇Cr-20的原生质体再生单核体为亲株,进行侧耳与香菇的属间原生质体融合,结果从1022个融合再生体中镜检出16个具锁状联合遗传标记的融合株。对1～4号融合株与亲株进行的拮抗反应、双单交配、过氧化物同功酶谱分析及菇形特征对比皆证明融合株确具双亲杂合体特征。出菇验证表明,融合株菌丝体长速均快于亲株,出菇期早,菇产量近于或高于亲株侧耳的水平。 相似文献