金针菇功能性蛋白的研究进展

主要综述了近年关于金针菇功能性蛋白,即核糖体失活蛋白、真菌免疫调节蛋白、金针菇毒素(一种成孔溶细胞素)的分离纯化及其性质,它们的生物学作用与作用机制和它们可能的应用及展望。     

金针菇多糖提取纯化研究进展

金针菇作为药食两用真菌,在我国的资源十分丰富.多糖是金针菇主要功能成分之一,具有抗氧化、抗衰老、抗肿瘤、抑菌、免疫调节和增加记忆等多种功能活性,研究金针菇多糖的提取纯化对金针菇综合开发应用具有重要意义.以近10年来国内发表文献(中国知网)为依据,总结概述金针菇多糖提取纯化研究进展,为金针菇多糖的深入研究及开发应用提供参...     

金针菇基因组测序及萜类合成关键基因分析

金针菇是我国一种重要的食药用真菌,具有较高的营养和药用价值,尤其是从金针菇中分离得到了多种麦角甾醇、倍半萜等萜类,具有抗菌、抗肿瘤和降血脂等功效。以往关于金针菇的研究多集中在营养成分、栽培和市场开发等方面,而很少报道有关生物活性成分的合成和代谢研究。从福建省金针菇主要栽培品种中获得原生质体单核化L11菌株,我们进行了金针菇基因组测序与初步分析,并利用生物信息学手段,研究了其主要的生物活性成分——萜类合成途径中的关键基因。结果显示,金针菇单孢全基因组序列长度为34.75Mb。通过分析萜类合成途径中的相关基因,确定了4个萜类合成关键基因的基因结构,并且表明其蛋白结构具有稳定性,该类基因还含有多个信号肽和跨膜结构。金针菇基因组测序的完成,为下一步功能基因的筛选、分析等研究工作奠定了基础。     

火菇素蛋白的免疫印迹检测*

从金针菇子实体浸提液中提取了抗癌活性物质火菇素,将纯化的火菇素用Freund佐剂乳化后注射到新西兰白兔体内,经数次加强免疫后采血并分离了抗血清,并以抗血清为探针建立了火菇素蛋白的免疫印迹法定性检测方法。应用免疫学方法检测金针菇菌丝体和子实体,结果显示,只在金针菇子实体中发现与火菇素抗体相结合的抗原信号。     

金针菇富锌条件及锌结合形态的研究

对金针菇深层发酵富锌培养条件进行了研究,并对Zn的生物有机化程度进行了初步探讨。结果表明,金针菇富锌的深层培养较好的Zn源为ZnAc_2,初始pH值为6.5,加入0.1％柠檬酸和0.4％的CMC-Na利于富锌和提高生物量。金针菇富锌过程中,93％以上的ZnAc_2以有机锌态存在,其中50％以上结合为Zn-蛋白,34％与糖及脂肪类物质结合。     

金针菇G蛋白偶联受体基因的CRISPR/Cas9基因组编辑载体构建及转化研究

通过氨基酸同源比对（Blast P）以及金针菇冷诱导前后菌丝阶段和原基阶段的转录组数据分析,获得了金针菇中的两个假定G蛋白偶联受体基因Fvgpcr1、Fvgpcr2。对获得的金针菇假定G蛋白偶联受体基因Fvgpcr1、Fvgpcr2构建了基因组编辑（CRISPR/Cas9）的pCAMBIA0390-hph-Fvcas9-Fvgpcr1- sgRNA1/sgRNA2、pCAMBIA0390-hph-Fvcas9-Fvgpcr2-sgRNA1/sgRNA2等4个表达载体。通过农杆菌介导（ATMT）将表达载体pCAMBIA0390-hph-Fvcas9-Fvgpcr-sgRNA转化金针菇菌丝体,采用潮霉素和头孢毒素低浓度初筛和高浓度复筛,经两段筛选获得金针菇拟转化子。经对拟转化子进行PCR鉴定、RT-qPCR检测和Western杂交验证,结果显示表达载体pCAMBIA0390-hph-Fvcas9-Fvgpcr-sgRNA成功整合进金针菇基因组中,FvCas9蛋白正常表达,但未得到Fvgpcr基因敲除突变体。本研究利用农杆菌介导转化法在金针菇中构建了CRISPR/Cas9敲除体系,对后续目标基因的敲除有着重要意义。     

基于金针菇全基因组的赖氨酸合成途径关键酶分析

【目的】以金针菇全基因组测序数据为基础研究其L-赖氨酸的从头合成途径及其关键基因。【方法】采用Illumina Hiseq2000和Roche454 FLX+两种方法完成金针菇单孢菌株Dan3的基因组测序,基于全基因组序列筛选金针菇中赖氨酸生物合成的关键基因,并分析这些基因编码的蛋白质基本理化性质;预测其亚细胞定位情况及蛋白的二级结构。【结果】金针菇Dan3全基因组序列长度为34.17 Mb,预测到8个参与α-氨基己二酸途径的关键基因,这些基因都含有多个内含子和外显子,二级结构主要由α-螺旋和无规则卷曲组成,有3个蛋白定位于线粒体。【结论】金针菇是通过α-氨基己二酸途径合成赖氨酸,基因组中预测到与该途径相关的几乎所有的酶。     

红油金针菇产品的辐照加工技术研究

研究了红油金针菇的辐照加工技术,通过正交试验确定了红油金针菇的最佳调味配方,结果表明：红油添加量10%、蔗糖添加量0.1%、食盐添加量3%、柠檬酸添加量0.2%为最佳调味配方。研究红油金针菇的防腐杀菌技术的结果表明：添加0.015%的脱氢乙酸钠和0.015%的山梨酸钾复配防腐剂比单一防腐剂更有效提高了红油金针菇的防腐效果,辐照剂量为4kGy杀菌后的红油金针菇可达到完全灭菌的效果。将上述杀菌处理后的红油金针菇进行常温贮藏,180d后红油金针菇产品细菌总数仍然很少并且保持有令人满意的色泽、脆度和口感。辐照处理对红油金针菇的营养成分影响不大。     

白色金针菇液体菌种制备及工厂化生产应用初探

通过对白色金针菇液体菌种制备及工厂化生产中应用的研究,确定白色金针菇液体菌种培养基配方及制备条件,建立了采用液体菌种进行白色金针菇工厂化生产的技术规程.通过对茵丝干重的测定,确定了液体菌种培养基配方、培养时间、培养温度、起始pH值等技术参数.并根据本地区的资源情况,通过比较试验,确定了白色金针菇液体菌种制备工艺,工厂化生产的培养料配方及工艺流程.结果表明:采用液体菌种进行白色金针菇的工厂化生产,发茵生长周期明显缩短,污染率显著降低,为白色金针菇工厂化生产提供了技术基础.     

金针菇成熟期和伸长期菌盖的转录组与蛋白组比较分析

菌盖是大型真菌的重要组成部分,也是其产生有性孢子的部位,但是其发育机制仍不明确。本研究以金针菇Flammulina filiformis为材料,采用转录组和蛋白组联合分析的方法,比较分析了金针菇成熟期和伸长期菌盖的差异基因与蛋白,并对其进行GO (gene ontology)功能聚类分析、KEGG (Kyoto encyclopedia of genes and genomes)富集分析和蛋白互作网络分析。本研究筛选到差异表达基因有1 391个,差异表达蛋白147个,均以上调表达为主。GO功能聚类分析结果表明,催化活性(catalytic activity)条目富集基因最多,其次是细胞组分(cell part)、细胞过程(cellular process)和细胞器(organelle)。KEGG富集分析结果表明,差异表达基因和蛋白主要富集在碳水化合物代谢通路(carbohydrate metabolism)和氨基酸代谢通路(amino acid metabolism)等。本研究选取了9个关键的差异表达基因,使用实时荧光定量PCR (real-time quantitative PCR,RT-qPCR)对其表达量进行了验证。RT-qPCR验证结果与转录组测序结果相一致。蛋白互作网络分析表明,水解酶类、结构域类和转录调节类蛋白为互作网络的主要结点。本研究联合转录组、蛋白组测序数据,通过分析差异基因与蛋白,为深入了解金针菇菌盖发育机制提供数据参考。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.