酿酒酵母缺陷型原生质体的形成再生及融合条件的探讨

营养缺陷型作为酿酒酵母(Saccharomyces Cerevisiae)单倍体融合亲株的遗传标记。采用营养缺陷型LY—2和LY—4菌侏的对数生长前期的细胞,在0.5％的蜗牛酶作用时间40分钟,0.7MKCl高渗稳定剂的条件下,制备原生质体。两亲株的原生质体形成率分别为98.2％和91.0％。原生质体再生率为6.12％和2.13％。两亲株的原生质体在35％聚乙二醇(PEG, M.W.6000),50mMCaCl_2下诱导融合15分钟,在基本培养基上长出营养互补的融合菌株,融合频率为4.14x10_(-4)。本文就两亲株的原生质体形成和再生条件进行了初步的探讨,并进行了原生质体融合的尝试,     

番茄果实乙烯释放量的遗传模型分析

本试验选择2个番茄果实乙烯释放量显著不同的番茄品系,通过P_1、P_2、F_1、F_2、B_1和B_2六世代的分析方法,研究了番茄果实乙烯释放量的遗传规律.结果表明:番茄果实乙烯释放量遗传符合1对负向完全显性主基因+加性-显性多基因模型(D-4),主基因效应在B_1、B_2和F_(23)个世代的遗传率分别为36.33%、44.09%和35.14%,多基因效应在B_1、B_2和F_(23)个世代的遗传率分别为54.73%、40.50%和54.88%.     

米曲霉尿嘧啶营养缺陷型突变体筛选及转化体系建立

米曲霉(Aspergillus oryzae)是一种十分重要的工业微生物，但由于其菌丝体和孢子均含有多个细胞核，并且对多种抗性药物具有抗性，导致其遗传转化和分子生物学研究较其他模式丝状真菌困难。目前米曲霉遗传转化主要采用营养缺陷型作为筛选标记。尿嘧啶营养缺陷型是米曲霉转化中最常用的一种营养缺陷型标记，其筛选标记基因pyrG编码乳清酸核苷-5′-磷酸脱羧酶为尿嘧啶的前体尿苷合成所必需。本研究以米曲霉3.042为背景，利用紫外线诱变和5-氟乳清酸(5-FOA)胁迫筛选获得5株尿嘧啶营养缺陷型菌株，经DNA测序5株菌株均为pyrG基因不同位点突变，确定为尿嘧啶营养缺陷型。以筛选到的尿嘧啶营养缺陷型菌株为背景，利用农杆菌介导的米曲霉转化系统，成功构建了米曲霉尿嘧啶营养缺陷型为筛选标记的农杆菌转化体系。     

宋内氏福氏2a双价志贺氏菌芳香族氨基酸营养缺陷型减毒株的研究:Ⅰ.减毒株的构建

FS54是本室构建的宋内氏福氏2a双价志贺氏菌杂交株,有与志贺氏菌毒株相同的毒力。本研究通过遗传重组技术,给FS54株引入了aroD基因的转座子Tn10插入失活物(aroD∶∶Tn10),构建了FS54株的芳香族氨基酸营养缺陷型减毒株,并进一步去除了Tn10所携带的四环素抗性和宋内氏Ⅰ相大质粒(pSS120∶∶Tn5)上Tn5所携带的卡那霉素抗性,得到了宋内氏福氏2a双价志贺氏菌芳香族氨基酸营养缺陷型减毒株FS54—7b。     

简讯

用曹养缺陷型的回复突变株提高谷氨酸生产菌的转化率用亚硝基胍处理谷氨酸生产菌钝齿棒状杆菌(Corynebacterium crenatum)B_9菌株,在100株营养缺陷型     

食源性沙门氏菌耐药性及质粒介导喹诺酮耐药基因检测

随机采集的638份食品样品中沙门氏菌总检出率为9.7%(n=62株),共检出16种不同的血清型,其中最常见的为鸭沙门氏菌。受试菌株对磺胺甲基异噁唑、复方新诺明、链霉素和环丙沙星的耐药率较高。16株耐环丙沙星沙门氏菌按GyrA和ParC喹诺酮耐药决定区(QRDR)不同氨基酸替代组合可分为5种突变型,其中GyrA亚基发生Ser83Phe和Asp87Gly变异,同时ParC亚基发生Ser80Arg变异为最常见的突变类型。62株食源性沙门氏菌中,qnr基因阳性的菌株共7株,占受试菌株的11.3%。qnrA和qnrS基因阳性菌株分别有2株和5株,没有菌株携带qnrB、qnrC和qnrD基因。aac(6')-Ib基因阳性菌株共有8株,其中3株经确认为携带其变体基因aac(6')-Ib-cr。结果表明,新乡市食源性沙门氏菌血清型分布呈多样性,耐药状况较为严重,并且一些菌株携带质粒介导喹诺酮耐药(PMQR)基因。     

利用可重复使用的URA3标记基因建立热带假丝酵母基因敲除系统

热带假丝酵母(Candida tropicalis)在发酵工业中具有重要的应用潜力,但二倍体遗传结构和较低的遗传转化效率限制了其代谢工程育种技术的应用。建立可靠的遗传转化技术并高效的删除目的基因是代谢工程改造热带假丝酵母的重要前提。文章以C. tropicalis ATCC 20336为出发菌株,通过化学诱变筛选获得了尿嘧啶缺陷型突变株C. tropicalis XZX(ura3/ura3)。以丙酮酸脱羧酶(Pyruvate decarboxylase,PDC)基因作为靶基因构建了两端包含同源臂并在选择性标记C. tropicalis URA3(Orotidine-5′-phosphate decarboxylase,乳清酸核苷-5-磷酸脱羧酶)基因两侧同向插入源于沙门氏菌(Salmonella typhimurium)的hisG序列的基因敲除盒PDC1-hisG-URA3-hisG- PDC1(PHUHP),并转化宿主菌株C. tropicalis XZX,筛选获得PHUHP片段正确整合到染色体的PDC基因位点的转化子XZX02。在此基础上,将转化子XZX02涂布于5-FOA(5-氟乳清酸)选择培养基上,筛选得到URA3基因从PHUHP片段中丢失的营养缺陷型菌株XZX03。进一步构建了第2个PDC等位基因的删除表达盒PDCm- URA3-PDCm,并转化C. tropicalis XZX03菌株,获得转化子C. tropicalis XZX04。经PCR和DNA测序确认转化子C. tropicalis XZX04细胞染色体上的两个PDC等位基因被成功敲除。文章建立了一种营养缺陷型标记可重复使用的热带假丝酵母遗传转化技术,利用该技术成功敲除了细胞的PDC基因,为进一步利用代谢工程改造热带假丝酵母奠定了基础。     

中国耐多药结核分枝杆菌临床分离株rpoB基因突变特点

为阐明中国耐多药结核分枝杆菌临床分离株rpoB基因的突变特征,对86株结核分枝杆菌临床分离株rpoB基因两个区域,包括81个碱基利福平抗药性决定区(rifampin resistance determining region,RRDR)和V176F区进行序列测定。其中72株耐多药分离株中的65株rpoB基因的RRDR区存在22种不同类型突变、21种点突变和一个插入突变。最常见的突变部位分别位于密码子531(41％)、526(40％)和516(4％),10％耐药分离株未检测到突变。鉴定了RRDR内6个新的等位基因,以及RRDR外部区域5个新的突变。所有分离菌株V176均无突变。     

耐甲氧西林金黄色葡萄球菌消毒剂和抗生素耐药相关基因的聚类分析

目的了解解放军第98医院临床分离的耐甲氧西林金黄色葡萄球菌(MRSA)抗菌药物耐药基因存在状况及菌株亲缘性。方法采用聚合酶链反应及序列分析的方法检测50株MRSA中10种耐药相关基因,采用Average法对耐药基因进行聚类分析。结果50株MRSA中m ecA、aac(6')-aph(2')、tetM和erm基因均阳性,qacA、blaTEM、aph(3')-III和ant(4',4')基因阳性率分别为92.0%、40.0%、98.0%和4.0%,vanA和vanB基因均阴性。在1号、2号、3号菌株的qacA基因序列编码区域同一位点均有1个碱基发生有义突变(G→A),相应的苏氨酸(T)被异亮氨酸(I)所取代。根据耐药基因的聚类分析该50株MRSA可分为2个亚群,为院内感染所致。结论临床分离的MRSA耐药相关基因携带率很高;qacA基因存在有义突变为新的发现;MRSA可导致克隆传播院内感染,并存在暴发性流行。     

利用营养缺陷诱变对金顶侧耳进行基因连锁分析研究

利用金顶侧耳的营养缺陷型菌株配制杂交菌株,通过营养缺陷型标记对其后代进行连锁分析,确定不亲和性因子和营养缺陷标记所在的连锁群及其排列顺序。截止目前的试验数据表明,金顶侧耳至少由6条染色体(连锁群)组成。其中A因子存在的第1连锁群上分布着ade2、pab1、ade5、met2、met7营养缺陷型基因位点;B(Bα、Bβ)因子存在的第2连锁群上分布着cho1、ade1营养缺陷型基因位点;第3连锁群上分布着met9、arg1、his1、ade7、his3、met1营养缺陷型基因位点;第4连锁群分布着nic1、nic2、ade4、pdx2营养缺陷型基因位点;第5连锁群分布着pan1、ino1营养缺陷型基因位点;第6连锁群分布着ile1营养缺陷型基因位点。金顶侧耳与其他担子菌的遗传图谱相同,第1连锁群A因子附近均分布着Ade及Pab营养缺陷型基因位点。     

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.